Journal articles on the topic 'Flagellati'
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1
Schuhmacher, Jan S., Florian Rossmann, Felix Dempwolff, Carina Knauer, Florian Altegoer, Wieland Steinchen, Anja K. Dörrich, et al. "MinD-like ATPase FlhG effects location and number of bacterial flagella during C-ring assembly." Proceedings of the National Academy of Sciences 112, no. 10 (March 2, 2015): 3092–97. http://dx.doi.org/10.1073/pnas.1419388112.
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The number and location of flagella, bacterial organelles of locomotion, are species specific and appear in regular patterns that represent one of the earliest taxonomic criteria in microbiology. However, the mechanisms that reproducibly establish these patterns during each round of cell division are poorly understood. FlhG (previously YlxH) is a major determinant for a variety of flagellation patterns. Here, we show that FlhG is a structural homolog of the ATPase MinD, which serves in cell-division site determination. Like MinD, FlhG forms homodimers that are dependent on ATP and lipids. It interacts with a complex of the flagellar C-ring proteins FliM and FliY (also FliN) in the Gram-positive, peritrichous-flagellated Bacillus subtilis and the Gram-negative, polar-flagellated Shewanella putrefaciens. FlhG interacts with FliM/FliY in a nucleotide-independent manner and activates FliM/FliY to assemble with the C-ring protein FliG in vitro. FlhG-driven assembly of the FliM/FliY/FliG complex is strongly enhanced by ATP and lipids. The protein shows a highly dynamic subcellular distribution between cytoplasm and flagellar basal bodies, suggesting that FlhG effects flagellar location and number during assembly of the C-ring. We describe the molecular evolution of a MinD-like ATPase into a flagellation pattern effector and suggest that the underappreciated structural diversity of the C-ring proteins might contribute to the formation of different flagellation patterns.
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Jensen, C., G. A. Schaub, and D. H. Molyneux. "The effect of Blastocrithidia triatomae (Trypanosomatidae) on the midgut of the reduviid bug Triatoma infestans." Parasitology 100, no. 1 (February 1990): 1–9. http://dx.doi.org/10.1017/s0031182000060054.
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SummaryThe pathogenic flagellate Blastocrithidia triatomae disrupts the digestion of Triatoma infestans; the midgut ultrastructure of bugs infected with the flagellate and of uninfected bugs is compared. Third or fourth instar larvae were dissected either unfed or 1 week after feeding.In all uninfected bugs extracellular membrane layers (e.m.l.) covered the apical microvillar border of the epithelial cells. Some midgut regions of bugs infected with B. triatomae appeared normal but often adjacent cells showed pathological effects. In affected cells the e.m.l. and the microvilli and finally the cells themselves were reduced or destroyed. Correlated with these observations of pathogenicity the method of attachment of parasites changed. When the e.m.l. were present only rarely were flagella found, but on extracellular membrane-free cells B. triatomae attached by flagellar enlargement to the microvillar border or, if this was reduced, to the apical host cell membrane. No hemidesmosome-like plaques were found at the attachment site. Although some flagella were inserted into the apical region of the cells no intracellular flagellates were observed.
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Nielsen, Lasse Tor, and Thomas Kiørboe. "Foraging trade-offs, flagellar arrangements, and flow architecture of planktonic protists." Proceedings of the National Academy of Sciences 118, no. 3 (January 11, 2021): e2009930118. http://dx.doi.org/10.1073/pnas.2009930118.
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Unicellular flagellated protists are a key element in aquatic microbial food webs. They all use flagella to swim and to generate feeding currents to encounter prey and enhance nutrient uptake. At the same time, the beating flagella create flow disturbances that attract flow-sensing predators. Protists have highly diverse flagellar arrangements in terms of number of flagella and their position, beat pattern, and kinematics, but it is unclear how the various arrangements optimize the fundamental trade-off between resource acquisition and predation risk. Here we describe the near-cell flow fields produced by 15 species and demonstrate consistent relationships between flagellar arrangement and swimming speed and between flagellar arrangement and flow architecture, and a trade-off between resource acquisition and predation risk. The flow fields fall in categories that are qualitatively described by simple point force models that include the drag force of the moving cell body and the propulsive forces of the flagella. The trade-off between resource acquisition and predation risk varies characteristically between flow architectures: Flagellates with multiple flagella have higher predation risk relative to their clearance rate compared to species with only one active flagellum, with the exception of the highly successful dinoflagellates that have simultaneously achieved high clearance rates and stealth behavior due to a unique flagellar arrangement. Microbial communities are shaped by trade-offs and environmental constraints, and a mechanistic explanation of foraging trade-offs is a vital part of understanding the eukaryotic communities that form the basis of pelagic food webs.
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Moestrup, Øjvind, and Jahn Throndsen. "Light and electron microscopical studies on Pseudoscourfieldia marina, a primitive scaly green flagellate (Prasinophyceae) with posterior flagella." Canadian Journal of Botany 66, no. 7 (July 1, 1988): 1415–34. http://dx.doi.org/10.1139/b88-197.
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The supposedly primitive prasinophyte Pseudoscourfieldia marina has been established in culture and examined in detail. The cells swim by means of two unequal but parallel posterior flagella. Pseudoscourfieldia differs from other green flagellates in the structure of the flagellar root system, possession of a conspicuous leaflike attenuated extension from one basal body into the cell, the structure of the pyrenoid, which is invaginated by long extensions from the mitochondrial complex, and the structure of the scaly covering on the body. Pseudoscourfieldia shows distinct similarities to Nephroselmis, which is laterally biflagellate, and to the Tetraselmis group, which possesses four anterior flagella. The attachment of the hair scales on the flagella has been examined in detail for the first time in a green flagellate. The very short hair scales attach to the flagellar surface between each pair of longitudinal rows of scales known as Melkonian's rows, associated with the inside of every third scale in one row. This is probably a feature common to all prasinophytes with Melkonian's rows. Based on the many investigations now published, a classification for the prasinophytes, supposedly some of the most primitive green algae, is suggested. One class, two orders, and four families are recognized and defined, with 15 genera.
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Wei, Xueming, and Wolfgang D. Bauer. "Starvation-Induced Changes in Motility, Chemotaxis, and Flagellation of Rhizobium meliloti." Applied and Environmental Microbiology 64, no. 5 (May 1, 1998): 1708–14. http://dx.doi.org/10.1128/aem.64.5.1708-1714.1998.
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ABSTRACT The changes in motility, chemotactic responsiveness, and flagellation of Rhizobium meliloti RMB7201, L5-30, and JJ1c10 were analyzed after transfer of the bacteria to buffer with no available C, N, or phosphate. Cells of these three strains remained viable for weeks after transfer to starvation buffer (SB) but lost all motility within just 8 to 72 h after transfer to SB. The rates of motility loss differed by severalfold among the strains. Each strain showed a transient, two- to sixfold increase in chemotactic responsiveness toward glutamine within a few hours after transfer to SB, even though motility dropped substantially during the same period. Strains L5-30 and JJ1c10 also showed increased responsiveness to the nonmetabolizable chemoattractant cycloleucine. Cycloleucine partially restored the motility of starving cells when added after transfer and prevented the loss of motility when included in the SB used for initial suspension of the cells. Thus, interactions between chemoattractants and their receptors appear to affect the regulation of motility in response to starvation independently of nutrient or energy source availability. Electron microscopic observations revealed that R. meliloti cells lost flagella and flagellar integrity during starvation, but not as fast, nor to such a great extent, as the cells lost motility. Even after prolonged starvation, when none of the cells were actively motile, about one-third to one-half of the initially flagellated cells retained some flagella. Inactivation of flagellar motors therefore appears to be a rapid and important response ofR. meliloti to starvation conditions. Flagellar-motor inactivation was at least partially reversible by addition of either cycloleucine or glucose. During starvation, some cells appeared to retain normal flagellation, normal motor activity, or both for relatively long periods while other cells rapidly lost flagella, motor activity, or both, indicating that starvation-induced regulation of motility may proceed differently in various cell subpopulations.
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Lowenthal, Andrew C., Marla Hill, Laura K. Sycuro, Khalid Mehmood, Nina R. Salama, and Karen M. Ottemann. "Functional Analysis of the Helicobacter pylori Flagellar Switch Proteins." Journal of Bacteriology 191, no. 23 (September 18, 2009): 7147–56. http://dx.doi.org/10.1128/jb.00749-09.
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ABSTRACT Helicobacter pylori uses flagellum-mediated chemotaxis to promote infection. Bacterial flagella change rotational direction by changing the state of the flagellar motor via a subcomplex referred to as the switch. Intriguingly, the H. pylori genome encodes four switch complex proteins, FliM, FliN, FliY, and FliG, instead of the more typical three of Escherichia coli or Bacillus subtilis. Our goal was to examine whether and how all four switch proteins participate in flagellation. Previous work determined that FliG was required for flagellation, and we extend those findings to show that all four switch proteins are necessary for normal numbers of flagellated cells. Furthermore, while fliY and fliN are partially redundant with each other, both are needed for wild-type levels of flagellation. We also report the isolation of an H. pylori strain containing an R54C substitution in fliM, resulting in bacteria that swim constantly and do not change direction. Along with data demonstrating that CheY-phosphate interacts with FliM, these findings suggest that FliM functions in H. pylori much as it does in other organisms.
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Murat, Dorothée, Marion Hérisse, Leon Espinosa, Alicia Bossa, François Alberto, and Long-Fei Wu. "Opposite and Coordinated Rotation of Amphitrichous Flagella Governs Oriented Swimming and Reversals in a Magnetotactic Spirillum." Journal of Bacteriology 197, no. 20 (August 3, 2015): 3275–82. http://dx.doi.org/10.1128/jb.00172-15.
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ABSTRACTCurrent knowledge regarding the mechanism that governs flagellar motor rotation in response to environmental stimuli stems mainly from the study of monotrichous and peritrichous bacteria. Little is known about how two polar flagella, one at each cell pole of the so-called amphitrichous bacterium, are coordinated to steer the swimming. Here we fluorescently labeled the flagella ofMagnetospirillum magneticumAMB-1 cells and took advantage of the magnetically controllable swimming of this bacterium to investigate flagellar rotation in moving cells. We identified three motility behaviors (runs, tumbles, and reversals) and two characteristic fluorescence patterns likely corresponding to flagella rotating in opposite directions. Each AMB-1 locomotion mode was systematically associated with particular flagellar patterns at the poles which led us to conclude that, while cell runs are allowed by the asymmetrical rotation of flagellar motors, their symmetrical rotation triggers cell tumbling. Our observations point toward a precise coordination of the two flagellar motors which can be temporarily unsynchronized during tumbling.IMPORTANCEMotility is essential for bacteria to search for optimal niches and survive. Many bacteria use one or several flagella to explore their environment. The mechanism by which bipolarly flagellated cells coordinate flagellar rotation is poorly understood. We took advantage of the genetic amenability and magnetically controlled swimming of the spirillum-shaped magnetotactic bacteriumMagnetospirillum magneticumAMB-1 to correlate cell motion with flagellar rotation. We found that asymmetric rotation of the flagella (counterclockwise at the lagging pole and clockwise at the leading pole) enables cell runs whereas symmetric rotation triggers cell tumbling. Taking into consideration similar observations in spirochetes, bacteria possessing bipolar ribbons of periplasmic flagella, we propose a conserved motility paradigm for spirillum-shaped bipolarly flagellated bacteria.
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Gray, Victoria L., Michael O'Reilly, Carsten T. Müller, Ian D. Watkins, and David Lloyd. "Low tyrosine content of growth media yields aflagellate Salmonella enterica serovar Typhimurium." Microbiology 152, no. 1 (January 1, 2006): 23–28. http://dx.doi.org/10.1099/mic.0.28442-0.
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Identification of Salmonella serotypes is based on flagellar and somatic antigens. The absence of flagella may consequently affect complete identification of the serotype; here it is shown that Salmonella enterica serovar Typhimurium exhibits morphological differences dependent on the peptone constituents of the culture medium. Aflagellate salmonella were produced in certain media where the nutritional ingredient was casein-based peptone or gelatin-based peptone; in gelatin-based peptone, aggregates of salmonella were observed. However, in media containing soy-based peptone as the primary nutrient, salmonella displayed a normal flagellated morphology. Transfer of aflagellate salmonella from nutritionally poor media, with casein- or gelatin-based peptone, into rich nutrient broth allowed flagella synthesis, indicating that the aflagellate form is still able to produce flagella. Amino acid sequencing of the peptones producing aflagellate organisms showed a relatively low tyrosine concentration: only 0·03±0·01 g l−1 for gelatin-based buffered peptone water, compared to 0·21±0·01 for soy-based buffered peptone water. Tyrosine is essential for flagellin, which is the subunit of the salmonella flagellar filament. The addition of 200 μM tyrosine to casein-based peptone media produced flagellate salmonella; 2 mM glucose was needed in addition to tyrosine to achieve a similar morphology in gelatin-based media. Therefore, culture media containing less than 1·20 g tyrosine l−1, and of limited carbohydrate source, when used for serological testing of clinical isolates, may result in an incomplete serological identification.
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ALLEN-VERCOE, E., A. R. SAYERS, and M. J. WOODWARD. "Virulence of Salmonella enterica serotype Enteritidis aflagellate and afimbriate mutants in a day-old chick model." Epidemiology and Infection 122, no. 3 (June 1999): 395–402. http://dx.doi.org/10.1017/s0950268899002460.
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Certain fimbriae and the flagellae of Salmonella enterica serovar Typhimurium have been shown to contribute to attachment and invasion of gut epithelium in the murine typhoid infection model and to contribute to pathogenesis in the chick. However, little is known of the role these organelles play in Enteritidis poultry infections and, to study this, day-old chicks were dosed orally in separate experiments with defined multiply afimbriate and/or aflagellate mutant strains of Enteritidis. The colonization and invasion characteristics of each mutant were compared with those of the isogenic wild type strain by the determination of the number of bacteria recovered from livers and spleens at known time points post infection. Compared with wild type Enteritidis, a mutant unable to express flagella but retaining the genetic potential to express fimbriae was recovered post mortem from livers and spleens in significantly reduced numbers compared to the isogenic wild-type at all time points post infection (P<0·001). Conversely, a flagellate but multiply afimbriate mutant (defective for the elaboration of five different fimbrial types) and a flagellate but non-motile ‘paralysed’ mutant were recovered from livers and spleens in similar numbers to the wild-type. The data suggested that Enteritidis flagella, but not fimbriae, played an important role in pathogenesis in the chick model and that the flagellar apparatus itself and not motility per se contributed significantly to this role.
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Bouteiller, Mathilde, Charly Dupont, Yvann Bourigault, Xavier Latour, Corinne Barbey, Yoan Konto-Ghiorghi, and Annabelle Merieau. "Pseudomonas Flagella: Generalities and Specificities." International Journal of Molecular Sciences 22, no. 7 (March 24, 2021): 3337. http://dx.doi.org/10.3390/ijms22073337.
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Flagella-driven motility is an important trait for bacterial colonization and virulence. Flagella rotate and propel bacteria in liquid or semi-liquid media to ensure such bacterial fitness. Bacterial flagella are composed of three parts: a membrane complex, a flexible-hook, and a flagellin filament. The most widely studied models in terms of the flagellar apparatus are E. coli and Salmonella. However, there are many differences between these enteric bacteria and the bacteria of the Pseudomonas genus. Enteric bacteria possess peritrichous flagella, in contrast to Pseudomonads, which possess polar flagella. In addition, flagellar gene expression in Pseudomonas is under a four-tiered regulatory circuit, whereas enteric bacteria express flagellar genes in a three-step manner. Here, we use knowledge of E. coli and Salmonella flagella to describe the general properties of flagella and then focus on the specificities of Pseudomonas flagella. After a description of flagellar structure, which is highly conserved among Gram-negative bacteria, we focus on the steps of flagellar assembly that differ between enteric and polar-flagellated bacteria. In addition, we summarize generalities concerning the fuel used for the production and rotation of the flagellar macromolecular complex. The last part summarizes known regulatory pathways and potential links with the type-six secretion system (T6SS).
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Jang, Moon-Sun, Yoshihiro Mouri, Kaoru Uchida, Shin-Ichi Aizawa, Masayuki Hayakawa, Nobuyuki Fujita, Takeaki Tezuka, and Yasuo Ohnishi. "Genetic and Transcriptional Analyses of the Flagellar Gene Cluster in Actinoplanes missouriensis." Journal of Bacteriology 198, no. 16 (June 6, 2016): 2219–27. http://dx.doi.org/10.1128/jb.00306-16.
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ABSTRACTActinoplanes missouriensis, a Gram-positive and soil-inhabiting bacterium, is a member of the rare actinomycetes. The filamentous cells produce sporangia, which contain hundreds of flagellated spores that can swim rapidly for a short period of time until they find niches for germination. These swimming cells are called zoospores, and the mechanism of this unique temporal flagellation has not been elucidated. Here, we report all of the flagellar genes in the bacterial genome and their expected function and contribution for flagellar morphogenesis. We identified a large flagellar gene cluster composed of 33 genes that encode the majority of proteins essential for assembling the functional flagella of Gram-positive bacteria. One noted exception to the cluster was the location of thefliQgene, which was separated from the cluster. We examined the involvement of four genes in flagellar biosynthesis by gene disruption,fliQ,fliC,fliK, andlytA. Furthermore, we performed a transcriptional analysis of the flagellar genes using RNA samples prepared fromA. missouriensisgrown on a sporangium-producing agar medium for 1, 3, 6, and 40 days. We demonstrated that the transcription of the flagellar genes was activated in conjunction with sporangium formation. Eleven transcriptional start points of the flagellar genes were determined using the rapid amplification of cDNA 5′ ends (RACE) procedure, which revealed the highly conserved promoter sequence CTCA(N15–17)GCCGAA. This result suggests that a sigma factor is responsible for the transcription of all flagellar genes and that the flagellar structure assembles simultaneously.IMPORTANCEThe biology of a zoospore is very interesting from the viewpoint of morphogenesis, survival strategy, and evolution. Here, we analyzed flagellar genes inA. missouriensis, which produces sporangia containing hundreds of flagellated spores each. Zoospores released from the sporangia swim for a short time before germination occurs. We identified a large flagellar gene cluster and an orphan flagellar gene (fliQ). These findings indicate that the zoospore flagellar components are typical of Gram-positive bacteria. However, the transcriptional analysis revealed that all flagellar genes are transcribed simultaneously during sporangium formation, a pattern differing from the orderly, regulated expression of flagellar genes in other bacteria, such asSalmonellaandEscherichia coli. These results suggest a novel regulatory mechanism for flagellar formation inA. missouriensis.
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Alexandre, Gladys, René Rohr, and René Bally. "A Phase Variant of Azospirillum lipoferum Lacks a Polar Flagellum and Constitutively Expresses Mechanosensing Lateral Flagella." Applied and Environmental Microbiology 65, no. 10 (October 1, 1999): 4701–4. http://dx.doi.org/10.1128/aem.65.10.4701-4704.1999.
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ABSTRACT Flagellation of a nonswimming variant of the mixed flagellated bacterium Azospirillum lipoferum 4B was characterized by electron microscopy, and polyclonal antibodies were raised against polar and lateral flagellins. The variant cells lacked a polar flagellum due to a defect in flagellin synthesis and constitutively expressed lateral flagella. The variant cells were able to respond to conditions that restricted the rotation of lateral flagella by producing more lateral flagella, suggesting that the lateral flagella, as well as the polar flagellum, are mechanosensing.
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Senesi, Sonia, Emilia Ghelardi, Francesco Celandroni, Sara Salvetti, Eva Parisio, and Alessandro Galizzi. "Surface-Associated Flagellum Formation and Swarming Differentiation in Bacillus subtilis Are Controlled by the ifm Locus." Journal of Bacteriology 186, no. 4 (February 15, 2004): 1158–64. http://dx.doi.org/10.1128/jb.186.4.1158-1164.2004.
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ABSTRACT Knowledge of the highly regulated processes governing the production of flagella in Bacillus subtilis is the result of several observations obtained from growing this microorganism in liquid cultures. No information is available regarding the regulation of flagellar formation in B. subtilis in response to contact with a solid surface. One of the best-characterized responses of flagellated eubacteria to surfaces is swarming motility, a coordinate cell differentiation process that allows collective movement of bacteria over solid substrates. This study describes the swarming ability of a B. subtilis hypermotile mutant harboring a mutation in the ifm locus that has long been known to affect the degree of flagellation and motility in liquid media. On solid media, the mutant produces elongated and hyperflagellated cells displaying a 10-fold increase in extracellular flagellin. In contrast to the mutant, the parental strain, as well as other laboratory strains carrying a wild-type ifm locus, fails to activate a swarm response. Furthermore, it stops to produce flagella when transferred from liquid to solid medium. Evidence is provided that the absence of flagella is due to the lack of flagellin gene expression. However, restoration of flagellin synthesis in cells overexpressing σD or carrying a deletion of flgM does not recover the ability to assemble flagella. Thus, the ifm gene plays a determinantal role in the ability of B. subtilis to contact with solid surfaces.
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Guo, Hanliang, Yi Man, Kirsty Y. Wan, and Eva Kanso. "Intracellular coupling modulates biflagellar synchrony." Journal of The Royal Society Interface 18, no. 174 (January 2021): 20200660. http://dx.doi.org/10.1098/rsif.2020.0660.
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Beating flagella exhibit a variety of synchronization modes. This synchrony has long been attributed to hydrodynamic coupling between the flagella. However, recent work with flagellated algae indicates that a mechanism internal to the cell, through the contractile fibres connecting the flagella basal bodies, must be at play to actively modulate flagellar synchrony. Exactly how basal coupling mediates flagellar coordination remains unclear. Here, we examine the role of basal coupling in the synchronization of the model biflagellate Chlamydomonas reinhardtii using a series of mathematical models of decreasing levels of complexity. We report that basal coupling is sufficient to achieve inphase, antiphase and bistable synchrony, even in the absence of hydrodynamic coupling and flagellar compliance. These modes can be reached by modulating the activity level of the individual flagella or the strength of the basal coupling. We observe a slip mode when allowing for differential flagellar activity, just as in experiments with live cells. We introduce a dimensionless ratio of flagellar activity to basal coupling that is predictive of the mode of synchrony. This ratio allows us to query biological parameters which are not yet directly measurable experimentally. Our work shows a concrete route for cells to actively control the synchronization of their flagella.
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Balaban, Murat, Stephanie N. Joslin, and David R. Hendrixson. "FlhF and Its GTPase Activity Are Required for Distinct Processes in Flagellar Gene Regulation and Biosynthesis in Campylobacter jejuni." Journal of Bacteriology 191, no. 21 (August 28, 2009): 6602–11. http://dx.doi.org/10.1128/jb.00884-09.
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ABSTRACT FlhF proteins are putative GTPases that are often necessary for one or more steps in flagellar organelle development in polarly flagellated bacteria. In Campylobacter jejuni, FlhF is required for σ54-dependent flagellar gene expression and flagellar biosynthesis, but how FlhF influences these processes is unknown. Furthermore, the GTPase activity of any FlhF protein and the requirement of this speculated activity for steps in flagellar biosynthesis remain uncharacterized. We show here that C. jejuni FlhF hydrolyzes GTP, indicating that these proteins are GTPases. C. jejuni mutants producing FlhF proteins with reduced GTPase activity were not severely defective for σ54-dependent flagellar gene expression, unlike a mutant lacking FlhF. Instead, these mutants had a propensity to lack flagella or produce flagella in improper numbers or at nonpolar locations, indicating that GTP hydrolysis by FlhF is required for proper flagellar biosynthesis. Additional studies focused on elucidating a possible role for FlhF in σ54-dependent flagellar gene expression were conducted. These studies revealed that FlhF does not influence production of or signaling between the flagellar export apparatus and the FlgSR two-component regulatory system to activate σ54. Instead, our data suggest that FlhF functions in an independent pathway that converges with or works downstream of the flagellar export apparatus-FlgSR pathway to influence σ54-dependent gene expression. This study provides corroborative biochemical and genetic analyses suggesting that different activities of the C. jejuni FlhF GTPase are required for distinct steps in flagellar gene expression and biosynthesis. Our findings are likely applicable to many polarly flagellated bacteria that utilize FlhF in flagellar biosynthesis processes.
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Tasteyre, Albert, Marie-Claude Barc, Anne Collignon, Helene Boureau, and Tuomo Karjalainen. "Role of FliC and FliD Flagellar Proteins ofClostridium difficile in Adherence and Gut Colonization." Infection and Immunity 69, no. 12 (December 1, 2001): 7937–40. http://dx.doi.org/10.1128/iai.69.12.7937-7940.2001.
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ABSTRACT In vitro and in vivo adhesive properties of flagella and recombinant flagellin FliC and flagellar cap FliD proteins ofClostridium difficile were analyzed. FliC, FliD, and crude flagella adhered in vitro to axenic mouse cecal mucus. Radiolabeled cultured cells bound to a high degree to FliD and weakly to flagella deposited on a membrane. The tissue association in the mouse cecum of a nonflagellated strain was 10-fold lower than that of a flagellated strain belonging to the same serogroup, confirming the role of flagella in adherence.
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Yamano, Kimiaki, Hiroshi Saito, Yukie Ogasawara, Shoko Fujii, Hiroko Yamada, Haruhisa Shirahama, and Hiroshi Kawai. "The Autofluorescent Substance in the Posterior Flagellum of Swarmers of the Brown Alga Scytosiphon lomentaria." Zeitschrift für Naturforschung C 51, no. 3-4 (April 1, 1996): 155–59. http://dx.doi.org/10.1515/znc-1996-3-404.
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Abstract The flagellar autofluorescent substance of the brown alga Scytosiphon lomentaria , which is probably involved in the photoreception of the phototaxis of flagellate cells, was investi gated. 4′,5′-Cyclic FMN (1) was isolated from the extract of whole mature plants for the first time as a natural product. Since the concentration of 4′,5′-cyclic FMN (1) was considerably low in vegetative plants, which do not contain fluorescent flagella, this substance is considered to correspond to the flagellar fluorescent substance.
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Roeßler, Markus, Gerhard Wanner, and Volker Müller. "Motility and Flagellum Synthesis inHalobacillus halophilus Are Chloride Dependent." Journal of Bacteriology 182, no. 2 (January 15, 2000): 532–35. http://dx.doi.org/10.1128/jb.182.2.532-535.2000.
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ABSTRACT The motility of Halobacillus halophilus as observed on swarm agar plates was strictly dependent on the chloride concentration. Cl− was apparently not used as the coupling ion for flagellar rotation. Cells grown in the absence of chloride were devoid of flagella, but flagellation was restored upon the addition of chloride. These experiments indicate that chloride is involved in synthesis of flagella in H. halophilus.
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Nakamura, Shuichi, and Tohru Minamino. "Flagella-Driven Motility of Bacteria." Biomolecules 9, no. 7 (July 14, 2019): 279. http://dx.doi.org/10.3390/biom9070279.
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The bacterial flagellum is a helical filamentous organelle responsible for motility. In bacterial species possessing flagella at the cell exterior, the long helical flagellar filament acts as a molecular screw to generate thrust. Meanwhile, the flagella of spirochetes reside within the periplasmic space and not only act as a cytoskeleton to determine the helicity of the cell body, but also rotate or undulate the helical cell body for propulsion. Despite structural diversity of the flagella among bacterial species, flagellated bacteria share a common rotary nanomachine, namely the flagellar motor, which is located at the base of the filament. The flagellar motor is composed of a rotor ring complex and multiple transmembrane stator units and converts the ion flux through an ion channel of each stator unit into the mechanical work required for motor rotation. Intracellular chemotactic signaling pathways regulate the direction of flagella-driven motility in response to changes in the environments, allowing bacteria to migrate towards more desirable environments for their survival. Recent experimental and theoretical studies have been deepening our understanding of the molecular mechanisms of the flagellar motor. In this review article, we describe the current understanding of the structure and dynamics of the bacterial flagellum.
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Portman, Neil, Christie Foster, Giselle Walker, and Jan Šlapeta. "Evidence of Intraflagellar Transport and Apical Complex Formation in a Free-Living Relative of the Apicomplexa." Eukaryotic Cell 13, no. 1 (September 20, 2013): 10–20. http://dx.doi.org/10.1128/ec.00155-13.
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ABSTRACT Since its first description, Chromera velia has attracted keen interest as the closest free-living relative of parasitic Apicomplexa. The life cycle of this unicellular alga is complex and involves a motile biflagellate form. Flagella are thought to be formed in the cytoplasm, a rare phenomenon shared with Plasmodium in which the canonical mode of flagellar assembly, intraflagellar transport, is dispensed with. Here we demonstrate the expression of intraflagellar transport components in C. velia , answering the question of whether this organism has the potential to assemble flagella via the canonical route. We have developed and characterized a culturing protocol that favors the generation of flagellate forms. From this, we have determined a marked shift in the mode of daughter cell production from two to four daughter cells per division as a function of time after passage. We conduct an ultrastructural examination of the C. velia flagellate form by using serial TEM and show that flagellar biogenesis in C. velia occurs prior to cytokinesis. We demonstrate a close association of the flagellar apparatus with a complex system of apical structures, including a micropore, a conoid, and a complex endomembrane system reminiscent of the apical complex of parasitic apicomplexans. Recent work has begun to elucidate the possible flagellar origins of the apical complex, and we show that in C. velia these structures are contemporaneous within a single cell and share multiple connections. We propose that C. velia therefore represents a vital piece in the puzzle of the origins of the apical complex.
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Blagotinsek, Vitan, Meike Schwan, Wieland Steinchen, Devid Mrusek, John C. Hook, Florian Rossmann, Sven A. Freibert, et al. "An ATP-dependent partner switch links flagellar C-ring assembly with gene expression." Proceedings of the National Academy of Sciences 117, no. 34 (August 11, 2020): 20826–35. http://dx.doi.org/10.1073/pnas.2006470117.
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Bacterial flagella differ in their number and spatial arrangement. In many species, the MinD-type ATPase FlhG (also YlxH/FleN) is central to the numerical control of bacterial flagella, and its deletion in polarly flagellated bacteria typically leads to hyperflagellation. The molecular mechanism underlying this numerical control, however, remains enigmatic. Using the model speciesShewanella putrefaciens, we show that FlhG links assembly of the flagellar C ring with the action of the master transcriptional regulator FlrA (named FleQ in other species). While FlrA and the flagellar C-ring protein FliM have an overlapping binding site on FlhG, their binding depends on the ATP-dependent dimerization state of FlhG. FliM interacts with FlhG independent of nucleotide binding, while FlrA exclusively interacts with the ATP-dependent FlhG dimer and stimulates FlhG ATPase activity. Our in vivo analysis of FlhG partner switching between FliM and FlrA reveals its mechanism in the numerical restriction of flagella, in which the transcriptional activity of FlrA is down-regulated through a negative feedback loop. Our study demonstrates another level of regulatory complexity underlying the spationumerical regulation of flagellar biogenesis and implies that flagellar assembly transcriptionally regulates the production of more initial building blocks.
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Lee, J. H., and C. J. Walsh. "Transcriptional regulation of coordinate changes in flagellar mRNAs during differentiation of Naegleria gruberi amebae into flagellates." Molecular and Cellular Biology 8, no. 6 (June 1988): 2280–87. http://dx.doi.org/10.1128/mcb.8.6.2280-2287.1988.
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The nuclear run-on technique was used to measure the rate of transcription of flagellar genes during the differentiation of Naegleria gruberi amebae into flagellates. Synthesis of mRNAs for the axonemal proteins alpha- and beta-tubulin and flagellar calmodulin, as well as a coordinately regulated poly(A)+ RNA that codes for an unidentified protein, showed transient increases averaging 22-fold. The rate of synthesis of two poly(A)+ RNAs common to amebae and flagellates was low until the transcription of the flagellar genes began to decline, at which time synthesis of the RNAs found in amebae increased 3- to 10-fold. The observed changes in the rate of transcription can account quantitatively for the 20-fold increase in flagellar mRNA concentration during the differentiation. The data for the flagellar calmodulin gene demonstrate transcriptional regulation for a nontubulin axonemal protein. The data also demonstrate at least two programs of transcriptional regulation during the differentiation and raise the intriguing possibility that some significant fraction of the nearly 200 different proteins of the flagellar axoneme is transcriptionally regulated during the 1 h it takes N. gruberi amebae to form visible flagella.
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Lee, J. H., and C. J. Walsh. "Transcriptional regulation of coordinate changes in flagellar mRNAs during differentiation of Naegleria gruberi amebae into flagellates." Molecular and Cellular Biology 8, no. 6 (June 1988): 2280–87. http://dx.doi.org/10.1128/mcb.8.6.2280.
Full textAbstract:
The nuclear run-on technique was used to measure the rate of transcription of flagellar genes during the differentiation of Naegleria gruberi amebae into flagellates. Synthesis of mRNAs for the axonemal proteins alpha- and beta-tubulin and flagellar calmodulin, as well as a coordinately regulated poly(A)+ RNA that codes for an unidentified protein, showed transient increases averaging 22-fold. The rate of synthesis of two poly(A)+ RNAs common to amebae and flagellates was low until the transcription of the flagellar genes began to decline, at which time synthesis of the RNAs found in amebae increased 3- to 10-fold. The observed changes in the rate of transcription can account quantitatively for the 20-fold increase in flagellar mRNA concentration during the differentiation. The data for the flagellar calmodulin gene demonstrate transcriptional regulation for a nontubulin axonemal protein. The data also demonstrate at least two programs of transcriptional regulation during the differentiation and raise the intriguing possibility that some significant fraction of the nearly 200 different proteins of the flagellar axoneme is transcriptionally regulated during the 1 h it takes N. gruberi amebae to form visible flagella.
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Lenaghan, Scott C., Stefan Nwandu-Vincent, Benjamin E. Reese, and Mingjun Zhang. "Unlocking the secrets of multi-flagellated propulsion: drawing insights from Tritrichomonas foetus." Journal of The Royal Society Interface 11, no. 93 (April 6, 2014): 20131149. http://dx.doi.org/10.1098/rsif.2013.1149.
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In this work, a high-speed imaging platform and a resistive force theory (RFT) based model were applied to investigate multi-flagellated propulsion, using Tritrichomonas foetus as an example. We discovered that T. foetus has distinct flagellar beating motions for linear swimming and turning, similar to the ‘run and tumble’ strategies observed in bacteria and Chlamydomonas . Quantitative analysis of the motion of each flagellum was achieved by determining the average flagella beat motion for both linear swimming and turning, and using the velocity of the flagella as inputs into the RFT model. The experimental approach was used to calculate the curvature along the length of the flagella throughout each stroke. It was found that the curvatures of the anterior flagella do not decrease monotonically along their lengths, confirming the ciliary waveform of these flagella. Further, the stiffness of the flagella was experimentally measured using nanoindentation, allowing for calculation of the flexural rigidity of T. foetus' s flagella, 1.55×10 −21 N m 2 . Finally, using the RFT model, it was discovered that the propulsive force of T. foetus was similar to that of sperm and Chlamydomonas , indicating that multi-flagellated propulsion does not necessarily contribute to greater thrust generation, and may have evolved for greater manoeuvrability or sensing. The results from this study have demonstrated the highly coordinated nature of multi-flagellated propulsion and have provided significant insights into the biology of T. foetus .
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Han, Ji Woong, Jong Ho Park, Misook Kim, and JooHun Lee. "mRNAs for Microtubule Proteins Are Specifically Colocalized during the Sequential Formation of Basal Body, Flagella, and Cytoskeletal Microtubules in the Differentiation of Naegleria gruberi." Journal of Cell Biology 137, no. 4 (May 19, 1997): 871–79. http://dx.doi.org/10.1083/jcb.137.4.871.
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We have examined the distribution of four mRNAs—α-tubulin, β-tubulin, flagellar calmodulin, and Class I mRNA—during differentiation of Naegleria gruberi amebas into flagellates by in situ hybridization. Three of the four mRNAs—α-tubulin, β-tubulin, and Class I mRNA—began to be colocalized at the periphery of the cells as soon as transcription of the respective genes was activated and before any microtubular structures were observable. At 70 min after the initiation of differentiation, these mRNAs were relocalized to the base of the growing flagella, adjacent to the basal bodies and microtubule organizing center for the cytoskeletal microtubules. Within an additional 15 min, the mRNAs were translocated to the posterior of the flagellated cells, and by the end of differentiation (120 min), very low levels of the mRNAs were observed. Cytochalasin D inhibited stage-specific localization of the mRNAs, demonstrating that RNA localization was actin dependent. Since cytochalasin D also blocked differentiation, this raises the possibility that actin-dependent RNA movement is an essential process for differentiation.
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Iino, T., Tomoko Oguchi, and T. Kuroiwa. "Polymorphism in a Flagellar-shape Mutant of Salmonella typhimurium." Microbiology 81, no. 1 (January 1, 2000): 37–45. http://dx.doi.org/10.1099/00221287-81-1-37.
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A flagellar-shape mutant, designated ‘polymorphous’, was isolated from a normal flagella strain of Salmonella typhimurium. The mutant produces normal flagella in phase 1 and polymorphous flagella in phase 2. The polymorphous flagella are either straight or possess one of the four distinct wave-forms, namely M, S, N or C, when observed with an electron microscope after negative staining with phosphotungstic acid or uranyl acetate. Conversions between the four wave-forms were found to be brought about mainly by a change in the degree of twisting of longitudinal strands around the axis of a flagella filament, without marked change in the relative lengths of the outermost and innermost strands. The major fraction of the polymorphous mutant flagella showed the N-form under any conditions of specimen preparation. The remaining four forms appeared as minor fractions in various proportions. Specimens fixed with formalin showed less pronounced polymorphism than unfixed ones. Negative staining with uranyl acetate was more effective than with phosphotungstic acid for observing polymorphism. Even though more than one form appeared among the polymorphous flagella, each individual flagellum comprised a single form except for a rare coexistence of S and N. The same form of flagella tended to coexist in a bacterium in a heteromorphously flagellated cell population. It was concluded that the conformation and arrangement of the flagellin molecules responsible for wave-form result from strong mutual interactions between the neighbouring molecules along the flagellar filaments and also, to a lesser extent, between the neighbouring filaments in a flagellar bundle, as well as being influenced by the physico-chemical environment.
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Marathe, Sandhya Amol, Arjun Balakrishnan, Vidya Devi Negi, Deepika Sakorey, Nagasuma Chandra, and Dipshikha Chakravortty. "Curcumin Reduces the Motility of Salmonella enterica Serovar Typhimurium by Binding to the Flagella, Thereby Leading to Flagellar Fragility and Shedding." Journal of Bacteriology 198, no. 13 (April 18, 2016): 1798–811. http://dx.doi.org/10.1128/jb.00092-16.
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ABSTRACTOne of the important virulence properties of the pathogen is its ability to travel to a favorable environment, cross the viscous mucus barrier (intestinal barrier for enteric pathogens), and reach the epithelia to initiate pathogenesis with the help of an appendage, like flagella. Nonetheless, flagella can act as an “Achilles heel,” revealing the pathogen's presence to the host through the stimulation of innate and adaptive immune responses. We assessed whether curcumin, a dietary polyphenol, could alter the motility ofSalmonella, a foodborne pathogen. It reduced the motility ofSalmonella entericaserovar Typhimurium by shortening the length of the flagellar filament (from ∼8 μm to ∼5 μm) and decreasing its density (4 or 5 flagella/bacterium instead of 8 or 9 flagella/bacterium). Upon curcumin treatment, the percentage of flagellated bacteria declined from ∼84% to 59%. However, no change was detected in the expression of the flagellin gene and protein. A fluorescence binding assay demonstrated binding of curcumin to the flagellar filament. This might make the filament fragile, breaking it into smaller fragments. Computational analysis predicted the binding of curcumin, its analogues, and its degraded products to a flagellin molecule at an interface between domains D1 and D2. Site-directed mutagenesis and a fluorescence binding assay confirmed the binding of curcumin to flagellin at residues ASN120, ASP123, ASN163, SER164, ASN173, and GLN175.IMPORTANCEThis work, to our knowledge the first report of its kind, examines how curcumin targets flagellar density and affects the pathogenesis of bacteria. We found that curcumin does not affect any of the flagellar synthesis genes. Instead, it binds to the flagellum and makes it fragile. It increases the torsional stress on the flagellar filament that then breaks, leaving fewer flagella around the bacteria. Flagella, which are crucial ligands for Toll-like receptor 5, are some of the most important appendages ofSalmonella. Curcumin is an important component of turmeric, which is a major spice used in Asian cooking. The loss of flagella can, in turn, change the pathogenesis of bacteria, making them more robust and fit in the host.
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Obuchowski, Pamela L., and Christine Jacobs-Wagner. "PflI, a Protein Involved in Flagellar Positioning in Caulobacter crescentus." Journal of Bacteriology 190, no. 5 (December 28, 2007): 1718–29. http://dx.doi.org/10.1128/jb.01706-07.
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ABSTRACT The bacterial flagellum is important for motility and adaptation to environmental niches. The sequence of events required for the synthesis of the flagellar apparatus has been extensively studied, yet the events that dictate where the flagellum is placed at the onset of flagellar biosynthesis remain largely unknown. We addressed this question for alphaproteobacteria by using the polarly flagellated alphaproteobacterium Caulobacter crescentus as an experimental model system. To identify candidates for a role in flagellar placement, we searched all available alphaproteobacterial genomes for genes of unknown function that cluster with early flagellar genes and that are present in polarly flagellated alphaproteobacteria while being absent in alphaproteobacteria with other flagellation patterns. From this in silico screen, we identified pflI. Loss of PflI function in C. crescentus results in an abnormally high frequency of cells with a randomly placed flagellum, while other aspects of cell polarization remain normal. In a wild-type background, a fusion of green fluorescent protein (GFP) and PflI localizes to the pole where the flagellum develops. This polar localization is independent of the flagellar protein FliF, whose oligomerization into the MS ring is thought to define the site of flagellar synthesis, suggesting that PflI acts before or independently of this event. Overproduction of PflI-GFP often leads to ectopic localization at the wrong, stalked pole. This is accompanied by a high frequency of flagellum formation at this ectopic site, suggesting that the location of PflI is a sufficient marker for a site for flagellar assembly.
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Wiese, Martin, Daniela Kuhn, and Christoph G. Grünfelder. "Protein Kinase Involved in Flagellar-Length Control." Eukaryotic Cell 2, no. 4 (August 2003): 769–77. http://dx.doi.org/10.1128/ec.2.4.769-777.2003.
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ABSTRACT During its life cycle, the parasitic protozoon Leishmania mexicana differentiates from a flagellated form, the promastigote, to an amastigote form carrying a rudimentary flagellum. Besides biochemical changes, this process involves a change in overall cell morphology including flagellar shortening. A mitogen-activated protein kinase kinase homologue designated LmxMKK was identified in a homology screening and found to be critically involved in the regulation of flagellar assembly and cell size. LmxMKK is exclusively expressed in the promastigote stage and is likely to be regulated by posttranslational mechanisms such as phosphorylation. A deletion mutant for the single-copy gene revealed motile flagella dramatically reduced in length and lacking the paraflagellar rod, a structure adjacent to the axoneme in kinetoplastid flagella. Moreover, a fraction of the cells showed perturbance of the axonemal structure. Complementation of the deletion mutant with the wild-type gene restored typical promastigote morphology. We propose that LmxMKK influences anterograde intraflagellar transport to maintain flagellar length in Leishmania promastigotes; as such, it is the first protein kinase known to be involved in organellar assembly.
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Karpov, Serguei A., and Alexander P. Mylnikov. "Ultrastructure of the colourless flagellate Hyperamoeba flagellata with special reference to the flagellar apparatus." European Journal of Protistology 33, no. 4 (December 1997): 349–55. http://dx.doi.org/10.1016/s0932-4739(97)80046-6.
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Diggins, M. A., and W. F. Dove. "Distribution of acetylated alpha-tubulin in Physarum polycephalum." Journal of Cell Biology 104, no. 2 (February 1, 1987): 303–9. http://dx.doi.org/10.1083/jcb.104.2.303.
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The expression and cytological distribution of acetylated alpha-tubulin was investigated in Physarum polycephalum. A monoclonal antibody specific for acetylated alpha-tubulin, 6-11B-1 (Piperno, G., and M. T. Fuller, 1985, J. Cell Biol., 101:2085-2094), was used to screen for this protein during three different stages of the Physarum life cycle--the amoeba, the flagellate, and the plasmodium. Western blots of two-dimensional gels of amoebal and flagellate proteins reveal that this antibody recognizes the alpha 3 tubulin isotype, which was previously shown to be formed by posttranslational modification (Green, L. L., and W. F. Dove, 1984, Mol. Cell. Biol., 4:1706-1711). Double-label immunofluorescence demonstrates that, in the flagellate, acetylated alpha-tubulin is localized in the flagella and flagellar cone. Similar experiments with amoebae interestingly reveal that only within the microtubule organizing center (MTOC) are there detectable amounts of acetylated alpha-tubulin. In contrast, the plasmodial stage gives no evidence for acetylated alpha-tubulin by Western blotting or by immunofluorescence.
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Yen, Jiun Y., Katherine M. Broadway, and Birgit E. Scharf. "Minimum Requirements of Flagellation and Motility for Infection of Agrobacterium sp. Strain H13-3 by Flagellotropic Bacteriophage 7-7-1." Applied and Environmental Microbiology 78, no. 20 (August 3, 2012): 7216–22. http://dx.doi.org/10.1128/aem.01082-12.
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ABSTRACTThe flagellotropic phage 7-7-1 specifically adsorbs toAgrobacteriumsp. strain H13-3 (formerlyRhizobium lupiniH13-3) flagella for efficient host infection. TheAgrobacteriumsp. H13-3 flagellum is complex and consists of three flagellin proteins: the primary flagellin FlaA, which is essential for motility, and the secondary flagellins FlaB and FlaD, which have minor functions in motility. Using quantitative infectivity assays, we showed that absence of FlaD had no effect on phage infection, while absence of FlaB resulted in a 2.5-fold increase in infectivity. AflaAdeletion strain, which produces straight and severely truncated flagella, experienced a significantly reduced infectivity, similar to that of aflaB flaDstrain, which produces a low number of straight flagella. A strain lacking all three flagellin genes is phage resistant. In addition to flagellation, flagellar rotation is required for infection. A strain that is nonmotile due to an in-frame deletion in the gene encoding the motor component MotA is resistant to phage infection. We also generated two strains with point mutations in themotAgene resulting in replacement of the conserved charged residue Glu98, which is important for modulation of rotary speed. A change to the neutral Gln caused the flagellar motor to rotate at a constant high speed, allowing a 2.2-fold-enhanced infectivity. A change to the positively charged Lys caused a jiggly motility phenotype with very slow flagellar rotation, which significantly reduced the efficiency of infection. In conclusion, flagellar number and length, as well as speed of flagellar rotation, are important determinants for infection by phage 7-7-1.
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Pion, Martin, Redouan Bshary, Saskia Bindschedler, Sevasti Filippidou, Lukas Y. Wick, Daniel Job, and Pilar Junier. "Gains of Bacterial Flagellar Motility in a Fungal World." Applied and Environmental Microbiology 79, no. 22 (August 30, 2013): 6862–67. http://dx.doi.org/10.1128/aem.01393-13.
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ABSTRACTThe maintenance of energetically costly flagella by bacteria in non-water-saturated media, such as soil, still presents an evolutionary conundrum. Potential explanations have focused on rare flooding events allowing dispersal. Such scenarios, however, overlook bacterial dispersal along mycelia as a possible transport mechanism in soils. The hypothesis tested in this study is that dispersal along fungal hyphae may lead to an increase in the fitness of flagellated bacteria and thus offer an alternative explanation for the maintenance of flagella even in unsaturated soils. Dispersal along fungal hyphae was shown for a diverse array of motile bacteria. To measure the fitness effect of dispersal, additional experiments were conducted in a model system mimicking limited dispersal, usingPseudomonas putidaKT2440 and its nonflagellated (ΔfliM) isogenic mutant in the absence or presence ofMorchella crassipesmycelia. In the absence of the fungus, flagellar motility was beneficial solely under conditions of water saturation allowing dispersal, while under conditions limiting dispersal, the nonflagellated mutant exhibited a higher level of fitness than the wild-type strain. In contrast, in the presence of a mycelial network under conditions limiting dispersal, the flagellated strain was able to disperse using the mycelial network and had a higher level of fitness than the mutant. On the basis of these results, we propose that the benefit of mycelium-associated dispersal helps explain the persistence of flagellar motility in non-water-saturated environments.
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Sampaio, Suely C. F., Wilson B. Luiz, Mônica A. M. Vieira, Rita C. C. Ferreira, Bruna G. Garcia, Rita Sinigaglia-Coimbra, Jorge L. M. Sampaio, Luís C. S. Ferreira, and Tânia A. T. Gomes. "Flagellar Cap Protein FliD Mediates Adherence of Atypical Enteropathogenic Escherichia coli to Enterocyte Microvilli." Infection and Immunity 84, no. 4 (February 1, 2016): 1112–22. http://dx.doi.org/10.1128/iai.01001-15.
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The expression of flagella correlates with different aspects of bacterial pathogenicity, ranging from adherence to host cells to activation of inflammatory responses by the innate immune system. In the present study, we investigated the role of flagella in the adherence of an atypical enteropathogenicEscherichia coli(aEPEC) strain (serotype O51:H40) to human enterocytes. Accordingly, isogenic mutants deficient in flagellin (FliC), the flagellar structural subunit; the flagellar cap protein (FliD); or the MotAB proteins, involved in the control of flagellar motion, were generated and tested for binding to differentiated Caco-2 cells. Binding of the aEPEC strain to enterocytes was significantly impaired in strains with thefliCandfliDgenes deleted, both of which could not form flagella on the bacterial surface. A nonmotile but flagellated MotAB mutant also showed impaired adhesion to Caco-2 cells. In accordance with these observations, adhesion of aEPEC strain 1711-4 to Caco-2 cells was drastically reduced after the treatment of Caco-2 cells with purified FliD. In addition, incubation of aEPEC bacteria with specific anti-FliD serum impaired binding to Caco-2 cells. Finally, incubation of Caco-2 cells with purified FliD, followed by immunolabeling, showed that the protein was specifically bound to the microvillus tips of differentiated Caco-2 cells. The aEPEC FliD or anti-FliD serum also reduced the adherence of prototype typical enteropathogenic, enterohemorrhagic, and enterotoxigenicE. colistrains to Caco-2 cells. In conclusion, our findings further strengthened the role of flagella in the adherence of aEPEC to human enterocytes and disclosed the relevant structural and functional involvement of FliD in the adhesion process.
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Raberg, Matthias, Frank Reinecke, Rudolf Reichelt, Ursula Malkus, Simone König, Markus Pötter, Wolfgang Florian Fricke, et al. "Ralstonia eutropha H16 Flagellation Changes According to Nutrient Supply and State of Poly(3-Hydroxybutyrate) Accumulation." Applied and Environmental Microbiology 74, no. 14 (May 23, 2008): 4477–90. http://dx.doi.org/10.1128/aem.00440-08.
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ABSTRACT Two-dimensional polyacrylamide gel electrophoresis (2D PAGE), in combination with matrix-assisted laser desorption ionization-time of flight analysis, and the recently revealed genome sequence of Ralstonia eutropha H16 were employed to detect and identify proteins that are differentially expressed during different phases of poly(3-hydroxybutyric acid) (PHB) metabolism. For this, a modified protein extraction protocol applicable to PHB-harboring cells was developed to enable 2D PAGE-based proteome analysis of such cells. Subsequently, samples from (i) the exponential growth phase, (ii) the stationary growth phase permissive for PHB biosynthesis, and (iii) a phase permissive for PHB mobilization were analyzed. Among several proteins exhibiting quantitative changes during the time course of a cultivation experiment, flagellin, which is the main protein of bacterial flagella, was identified. Initial investigations that report on changes of flagellation for R. eutropha were done, but 2D PAGE and electron microscopic examinations of cells revealed clear evidence that R. eutropha exhibited further significant changes in flagellation depending on the life cycle, nutritional supply, and, in particular, PHB metabolism. The results of our study suggest that R. eutropha is strongly flagellated in the exponential growth phase and loses a certain number of flagella in transition to the stationary phase. In the stationary phase under conditions permissive for PHB biosynthesis, flagellation of cells admittedly stagnated. However, under conditions permissive for intracellular PHB mobilization after a nitrogen source was added to cells that are carbon deprived but with full PHB accumulation, flagella are lost. This might be due to a degradation of flagella; at least, the cells stopped flagellin synthesis while normal degradation continued. In contrast, under nutrient limitation or the loss of phasins, cells retained their flagella.
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Kühn, Marco J., Felix K. Schmidt, Bruno Eckhardt, and Kai M. Thormann. "Bacteria exploit a polymorphic instability of the flagellar filament to escape from traps." Proceedings of the National Academy of Sciences 114, no. 24 (May 30, 2017): 6340–45. http://dx.doi.org/10.1073/pnas.1701644114.
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Many bacterial species swim by rotating single polar helical flagella. Depending on the direction of rotation, they can swim forward or backward and change directions to move along chemical gradients but also to navigate their obstructed natural environment in soils, sediments, or mucus. When they get stuck, they naturally try to back out, but they can also resort to a radically different flagellar mode, which we discovered here. Using high-speed microscopy, we monitored the swimming behavior of the monopolarly flagellated speciesShewanella putrefacienswith fluorescently labeled flagellar filaments at an agarose–glass interface. We show that, when a cell gets stuck, the polar flagellar filament executes a polymorphic change into a spiral-like form that wraps around the cell body in a spiral-like fashion and enables the cell to escape by a screw-like backward motion. Microscopy and modeling suggest that this propagation mode is triggered by an instability of the flagellum under reversal of the rotation and the applied torque. The switch is reversible and bacteria that have escaped the trap can return to their normal swimming mode by another reversal of motor direction. The screw-type flagellar arrangement enables a unique mode of propagation and, given the large number of polarly flagellated bacteria, we expect it to be a common and widespread escape or motility mode in complex and structured environments.
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Drake, David, and Thomas C. Montie. "Protection against Pseudomonas aeruginosa infection by passive transfer of anti-flagellar serum." Canadian Journal of Microbiology 33, no. 9 (September 1, 1987): 755–63. http://dx.doi.org/10.1139/m87-130.
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The specificity of adsorbed flagellar antisera for H-antigen was demonstrated in vitro by cross-agglutination assays, motility inhibition, and an ELISA. The specific flagellar antibody was determined to be an IgG. Complete protection against burn wound sepsis was achieved with flagellar antisera. Cross-protection experiments revealed that protection was not only H-antigen dependent, but specific for the flagella antigen type. Antiserum raised against b-type flagella would only protect against homologous bacterial challenge and not against a-type flagellated strains. Results using a-type antisera were consistent, giving protection only against the homologous strain. In contrast, protective capacity was selectively removed from antisera by adsorbing with Fla+ cells. Bacteria colonized the burn wounds of passively protected mice to similar levels as seen in nonprotected animals, but the colonization remained localized and did not result in systemic infection, a pattern similar to infections with motility mutants observed in other studies. Animals rendered neutropenic prior to burning were not protected with flagellar antisera. These data suggested a role for phagocytic cells in protection. Immobilization by flagellar antiserum was observed both by microscopic studies and by inhibition of colony spreading. Antiflagellar antibody is hypothesized as exerting its protective capacity possibly in two ways; first by inhibiting the motility of invading bacteria by binding to the flagellum and immobilizing the bacteria, and secondly by acting as an opsonin, targeting either immobilized or mobile cells for phagocytosis.
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McFadden, G. I., D. Schulze, B. Surek, J. L. Salisbury, and M. Melkonian. "Basal body reorientation mediated by a Ca2+-modulated contractile protein." Journal of Cell Biology 105, no. 2 (August 1, 1987): 903–12. http://dx.doi.org/10.1083/jcb.105.2.903.
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A rapid, Ca2+-dependent change in the angle between basal bodies (up to 180 degrees) is associated with light-induced reversal of swimming direction (the "photophobic" response) in a number of flagellated green algae. In isolated, detergent-extracted, reactivated flagellar apparatus complexes of Spermatozopsis similis, axonemal beat form conversion to the symmetrical/undulating flagellar pattern and basal body reorientation (from the antiparallel to the parallel configuration) are simultaneously induced at greater than or equal to 10(-7) M Ca2+. Basal body reorientation, however, is independent of flagellar beating since it is induced at greater than or equal to 10(-7) M Ca2+ when flagellar beating is inhibited (i.e., in the presence of 1 microM orthovanadate in reactivation solutions; in the absence of ATP or dithiothreitol in isolation and reactivation solutions), or when axonemes are mechanically removed from flagellar apparatuses. Although frequent axonemal beat form reversals were induced by varying the Ca2+ concentration, antiparallel basal body configuration could not be restored in isolated flagellar apparatuses. Observations of the photophobic response in vivo indicate that even though the flagella resume the asymmetric, breaststroke beat form 1-2 s after photostimulation, antiparallel basal body configuration is not restored until a few minutes later. Using an antibody generated against the 20-kD Ca2+-modulated contractile protein of striated flagellar roots of Tetraselmis striata (Salisbury, J. L., A. Baron, B. Surek, and M. Melkonian, 1984, J. Cell Biol., 99:962-970), we have found the distal connecting fiber of Spermatozopsis similis to be immunoreactive by indirect immunofluorescence and immunogold electron microscopy. Electrophoretic and immunoblot analysis indicates that the antigen of S. similis flagellar apparatuses consists, like the Tetraselmis protein, of two acidic isoforms of 20 kD. We conclude that the distal basal body connecting fiber is a contractile organelle and reorients basal bodies during the photophobic response in certain flagellated green algae.
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Newell, D. G. "Monoclonal antibodies directed against the flagella ofCampylobacter jejuni: cross-reacting and serotypic specificity and potential use in diagnosis." Journal of Hygiene 96, no. 3 (June 1986): 377–84. http://dx.doi.org/10.1017/s0022172400066134.
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SUMMARYNine monoclonal antibodies directed against the flagella ofCampylobacter jejunistrain 81116 have been investigated for serotypic and cross-reacting activity using a panel of 17 Penner serotype strains ofC. jejuni.Four monoclonal antibodies were exclusively specific for serotype-6 strains, which was the serotype ofC. jejunistrain 81116. Two monoclonal antibodies cross-reacted with all the flagellated strains ofC jejunitested. One of these cross-reacting monoclonal antibodies, CF5, was found to react with all otherCampylobacterspecies exceptC. sputorum bulbulusbut it did not react with other bacterial entcropathogens. An antigencapture ELISA technique was established, using this monoclonal antibody, which could detect flagellar antigen in human faecal material. These anti-flagella monoclonal antibodies therefore may be valuable in the diagnosis and serotyping ofC. jejuniin clinical material.
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Konishi, Manabu, Masaomi Kanbe, Jonathan L. McMurry, and Shin-Ichi Aizawa. "Flagellar Formation in C-Ring-Defective Mutants by Overproduction of FliI, the ATPase Specific for Flagellar Type III Secretion." Journal of Bacteriology 191, no. 19 (July 31, 2009): 6186–91. http://dx.doi.org/10.1128/jb.00601-09.
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ABSTRACT The flagellar cytoplasmic ring (C ring), which consists of three proteins, FliG, FliM, and FliN, is located on the cytoplasmic side of the flagellum. The C ring is a multifunctional structure necessary for flagellar protein secretion, torque generation, and switching of the rotational direction of the motor. The deletion of any one of the fliG, fliM, and fliN genes results in a Fla− phenotype. Here, we show that the overproduction of the flagellum-specific ATPase FliI overcomes the inability of basal bodies with partial C-ring structures to produce complete flagella. Flagella made upon FliI overproduction were paralyzed, indicating that an intact C ring is essential for motor function. In FliN- or FliM-deficient mutants, flagellum production was about 10% of the wild-type level, while it was only a few percent in FliG-deficient mutants, suggesting that the size of partial C rings affects the extent of flagellation. For flagella made in C-ring mutants, the hook length varied considerably, with many being markedly shorter or longer than that of the wild type. The broad distribution of hook lengths suggests that defective C rings cannot control the hook length as tightly as the wild type even though FliK and FlhB are both intact.
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Thomsen, Helge A., Kurt R. Buck, Patricia A. Bolt, and David L. Garrison. "Fine structure and biology of Cryothecomonas gen. nov. (Protista incertae sedis) from the ice biota." Canadian Journal of Zoology 69, no. 4 (April 1, 1991): 1048–70. http://dx.doi.org/10.1139/z91-150.
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The morphology and ultrastructure of four species of Cryothecomonas gen.nov. (Protista incertae sedis) in material from the Weddell Sea, Antarctica, and the Isefjord, Denmark, are described. These heterotrophic flagellates, which were initially observed in association with sea ice, display a unique combination of morphological characteristics. At present it is impossible to assign the new genus to an existing higher taxonomic level of protistan flagellates. Cryothecomonas species are furnished with a close-fitting multilayered theca. The two naked anterior flagella emerge through narrow thecal funnels. A transitional helix is part of the flagellar transition zone. A conspicuous cytostome is located in a posterior (lateral) position. Food uptake is mediated through the extension of cytostomal pseudopodia. The nucleus is anteriorly located and contains a conspicuous nucleolus and distinct areas of chromatin. Mitochondrial cristae are tubular. Cryothecomonas species feed on cells in the size range 2–4.5 μm (e.g., algal flagellates). Data are presented on the abundance of Cryothecomonas armigera sp.nov. in Antarctic waters.
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Newell, Diane G., Harold McBride, and Jean M. Dolby. "Investigations on the role of flagella in the colonization of infant mice withCampylobacter jejuniand attachment ofCampylobacter jejunito human epithelial cell lines." Journal of Hygiene 95, no. 2 (October 1985): 217–27. http://dx.doi.org/10.1017/s0022172400062653.
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SUMMARYThe biochemical and biological properties of the flagella ofCampylobacter jejunihave been investigated using two variants selected from a flagellate, motile clinical isolate (strain 81116): a flagellate, non-motile variant (SF-1) and an aflagellate variant (SF-2). Phenotypic and biochemical analysis of the strains and amino acid analysis of the isolated flagella suggest that the variants differed from the wild-type strain only in the absence of flagella and/or motility. The aflagellato variant poorly colonized the gastrointestinal tract of infant mice but the flagellate, non-motile variant colonized the mice as successfully as the wild-type strain.35S-labelled organisms were used to investigate the attachment of the variants to human epithelial cell monolayersin vitro. The flagellate, non-motile strain attached more efficiently to the cells than the wild-type strain or the aflagellate strain. Differences in attachment suggest that an adhesin is intimately associated with flagella ofC jejuniand that active flagella mediate only a tenuous association with host cells. This adhesin attached most efficiently to cells of intestinal epithelial origin and was not specifically inhibited by various sugars.
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Soscia, Chantal, Abderrahman Hachani, Alain Bernadac, Alain Filloux, and Sophie Bleves. "Cross Talk between Type III Secretion and Flagellar Assembly Systems in Pseudomonas aeruginosa." Journal of Bacteriology 189, no. 8 (February 16, 2007): 3124–32. http://dx.doi.org/10.1128/jb.01677-06.
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ABSTRACT Pseudomonas aeruginosa cytotoxicity is linked to a type III secretion system (T3SS) that delivers effectors into the host cell. We show here that a negative cross-control exists between T3SS and flagellar assembly. We observed that, in a strain lacking flagella, T3SS gene expression, effector secretion, and cytotoxicity were increased. Conversely, we revealed that flagellar-gene expression and motility were decreased in a strain overproducing ExsA, the T3SS master regulator. Interestingly, a nonmotile strain lacking the flagellar filament (ΔfliC) presented a hyperefficient T3SS and a nonmotile strain assembling flagella (ΔmotAB) did not. More intriguingly, a strain lacking motCD genes is a flagellated strain with a slight defect in swimming. However, in this strain, T3SS gene expression was up-regulated. These results suggest that flagellar assembly and/or mobility antagonizes the T3SS and that a negative cross talk exists between these two systems. An illustration of this is the visualization by electron microscopy of T3SS needles in a nonmotile P. aeruginosa strain, needles which otherwise are not detected. The molecular basis of the cross talk is complex and remains to be elucidated, but proteins like MotCD might have a crucial role in signaling between the two processes. In addition, we found that the GacA response regulator negatively affects the T3SS. In a gacA mutant, the T3SS effector ExoS is hypersecreted. Strikingly, GacA was previously reported as a positive regulator for motility. Globally, our data document the idea that some virulence factors are coordinately but inversely regulated, depending on the bacterial colonization phase and infection types.
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Liu, Yujie, Rory Claydon, Marco Polin, and Douglas R. Brumley. "Transitions in synchronization states of model cilia through basal-connection coupling." Journal of The Royal Society Interface 15, no. 147 (October 2018): 20180450. http://dx.doi.org/10.1098/rsif.2018.0450.
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Despite evidence for a hydrodynamic origin of flagellar synchronization between different eukaryotic cells, recent experiments have shown that in single multi-flagellated organisms, coordination hinges instead on direct basal body connections. The mechanism by which these connections lead to coordination, however, is currently not understood. Here, we focus on the model biflagellate Chlamydomonas reinhardtii , and propose a minimal model for the synchronization of its two flagella as a result of both hydrodynamic and direct mechanical coupling. A spectrum of different types of coordination can be selected, depending on small changes in the stiffness of intracellular couplings. These include prolonged in-phase and anti-phase synchronization, as well as a range of multi-stable states induced by spontaneous symmetry breaking of the system. Linking synchrony to intracellular stiffness could lead to the use of flagellar dynamics as a probe for the mechanical state of the cell.
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Houry, A., R. Briandet, S. Aymerich, and M. Gohar. "Involvement of motility and flagella in Bacillus cereus biofilm formation." Microbiology 156, no. 4 (April 1, 2010): 1009–18. http://dx.doi.org/10.1099/mic.0.034827-0.
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Bacillus cereus is a food-borne pathogen and a frequent contaminant of food production plants. The persistence of this pathogen in various environments results from the formation of spores and of biofilms. To investigate the role of the B. cereus flagellar apparatus in biofilm formation, we constructed a non-flagellated mutant and a flagellated but non-motile mutant. Unexpectedly, we found that the presence of flagella decreased the adhesion of the bacterium to glass surfaces. We hypothesize that this decrease is a consequence of the flagella hindering a direct interaction between the bacterial cell wall and the surface. In contrast, in specific conditions, motility promotes biofilm formation. Our results suggest that motility could influence biofilm formation by three mechanisms. Motility is necessary for the bacteria to reach surfaces suitable for biofilm formation. In static conditions, reaching the air–liquid interface, where the biofilm forms, is a strong requirement, whereas in flow cells bacteria can have access to the bottom glass slide by sedimentation. Therefore, motility is important for biofilm formation in glass tubes and in microtitre plates, but not in flow cells. Motility also promotes recruitment of planktonic cells within the biofilm by allowing motile bacteria to invade the whole biofilm. Finally, motility is involved in the spreading of the biofilm on glass surfaces.
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Filip’echeva, Yulia A., Andrei V. Shelud’ko, Alexei G. Prilipov, Gennady L. Burygin, Elizaveta M. Telesheva, Stella S. Yevstigneyeva, Marina P. Chernyshova, Lilia P. Petrova, and Elena I. Katsy. "Plasmid AZOBR_p1-borne fabG gene for putative 3-oxoacyl-[acyl-carrier protein] reductase is essential for proper assembly and work of the dual flagellar system in the alphaproteobacterium Azospirillum brasilense Sp245." Canadian Journal of Microbiology 64, no. 2 (February 2018): 107–18. http://dx.doi.org/10.1139/cjm-2017-0561.
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Azospirillum brasilense can swim and swarm owing to the activity of a constitutive polar flagellum (Fla) and inducible lateral flagella (Laf), respectively. Experimental data on the regulation of the Fla and Laf assembly in azospirilla are scarce. Here, the coding sequence (CDS) AZOBR_p1160043 (fabG1) for a putative 3-oxoacyl-[acyl-carrier protein (ACP)] reductase was found essential for the construction of both types of flagella. In an immotile leaky Fla− Laf− fabG1::Omegon-Km mutant, Sp245.1610, defects in flagellation and motility were fully complemented by expressing the CDS AZOBR_p1160043 from plasmid pRK415. When pRK415 with the cloned CDS AZOBR_p1160045 (fliC) for a putative 65.2 kDa Sp245 Fla flagellin was transferred into the Sp245.1610 cells, the bacteria also became able to assemble a motile single flagellum. Some cells, however, had unusual swimming behavior, probably because of the side location of the organelle. Although the assembly of Laf was not restored in Sp245.1610 (pRK415-p1160045), this strain was somewhat capable of swarming motility. We propose that the putative 3-oxoacyl-[ACP] reductase encoded by the CDS AZOBR_p1160043 plays a role in correct flagellar location in the cell envelope and (or) in flagellar modification(s), which are also required for the inducible construction of Laf and for proper swimming and swarming motility of A. brasilense Sp245.
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O'Neil, Heather S., and Hélène Marquis. "Listeria monocytogenes Flagella Are Used for Motility, Not as Adhesins, To Increase Host Cell Invasion." Infection and Immunity 74, no. 12 (September 18, 2006): 6675–81. http://dx.doi.org/10.1128/iai.00886-06.
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ABSTRACT Flagellar structures contribute to the virulence of multiple gastrointestinal pathogens either as the effectors of motility, as adhesins, or as a secretion apparatus for virulence factors. Listeria monocytogenes is a food-borne, gram-positive pathogen that uses flagella to increase the efficiency of epithelial cell invasion (A. Bigot, H. Pagniez, E. Botton, C. Frehel, I. Dubail, C. Jacquet, A. Charbit, and C. Raynaud, Infect. Immun. 73:5530-5539, 2005; L. Dons, E. Eriksson, Y. Jin, M. E. Rottenberg, K. Kristensson, C. N. Larsen, J. Bresciani, and J. E. Olsen, Infect. Immun. 72:3237-3244, 2004). In this study, we aimed to elucidate the mechanism by which flagella contribute to L. monocytogenes invasion. To examine the role of flagella as adhesins, invasion and adhesion assays were performed with flagellated motile and nonmotile bacteria and nonflagellated bacteria. We observed that flagellated but nonmotile bacteria do not adhere to or invade human epithelial cells more efficiently than nonflagellated bacteria. These results indicated that flagella do not function as adhesins to enhance the adhesion of L. monocytogenes to targeted host cells. Instead, it appears that motility is important for tissue culture invasion. Furthermore, we tested whether motility contributes to early colonization of the gastrointestinal tract using a competitive index assay in which mice were infected orally with motile and nonmotile bacteria in a 1:1 ratio. Differential bacterial counts demonstrated that motile bacteria outcompete nonmotile bacteria in the colonization of the intestines at early time points postinfection. This difference is also reflected in invasion of the liver 12 h later, suggesting that flagellum-mediated motility enhances L. monocytogenes infectivity soon after bacterial ingestion in vivo.
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Dentler, WL, and C. Adams. "Flagellar microtubule dynamics in Chlamydomonas: cytochalasin D induces periods of microtubule shortening and elongation; and colchicine induces disassembly of the distal, but not proximal, half of the flagellum." Journal of Cell Biology 117, no. 6 (June 15, 1992): 1289–98. http://dx.doi.org/10.1083/jcb.117.6.1289.
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To study the mechanisms responsible for the regulation of flagellar length, we examined the effects of colchicine and Cytochalasin D (CD) on the growth and maintenance of Chlamydomonas flagella on motile wild type cells as well as on pf 18 cells, whose flagella lack the central microtubules and are immobile. CD had no effect on the regeneration of flagella after deflagellation but it induced fully assembled flagella to shorten at an average rate of 0.03 microns-min. Cells remained fully motile in CD and even stubby flagella continued to move, indicating that flagellar shortening did not selectively disrupt machinery necessary for motility. To observe the effects of the drug on individual cells, pf 18 cells were treated with CD and flagella on cells were monitored by direct observation over a 5-hour period. Flagella on control pf 18 cells maintained their initial lengths throughout the experiment but flagella on CD-treated cells exhibited periods of elongation, shortening, and regrowth suggestive of the dynamic behavior of cytoplasmic microtubules observed in vitro and in vitro. Cells behaved individually, with no two cells exhibiting the same flagellar behavior at any given time although both flagella on any single cell behaved identically. The rate of drug-induced flagellar shortening and elongation in pf 18 cells varied from 0.08 to 0.17 microns-min-1, with each event occurring over 10-60-min periods. Addition of colchicine to wild type and pf 18 cells induced flagella to shorten at an average rate of 0.06 microns-min-1 until the flagella reached an average of 73% of their initial length, after which they exhibited no further shortening or elongation. Cells treated with colchicine and CD exhibited nearly complete flagellar resorption, with little variation in flagellar length among cells. The effects of these drugs were reversible and flagella grew to normal stable lengths after drug removal. Taken together, these results show that the distal half to one-third of the Chlamydomonas flagellum is relatively unstable in the presence of colchicine but that the proximal half to two-thirds of the flagellum is stable to this drug. In contrast to colchicine, CD can induce nearly complete flagellar microtubule disassembly as well as flagellar assembly. Flagellar microtubules must, therefore, be inherently unstable, and flagellar length is stabilized by factors that are sensitive, either directly or indirectly, to the effects of CD.
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Fulano, Alex M., Danyu Shen, Miki Kinoshita, Shan-Ho Chou, and Guoliang Qian. "The Homologous Components of Flagellar Type III Protein Apparatus Have Acquired a Novel Function to Control Twitching Motility in a Non-Flagellated Biocontrol Bacterium." Biomolecules 10, no. 5 (May 7, 2020): 733. http://dx.doi.org/10.3390/biom10050733.
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The bacterial flagellum is one of the best-studied surface-attached appendages in bacteria. Flagellar assembly in vivo is promoted by its own protein export apparatus, a type III secretion system (T3SS) in pathogenic bacteria. Lysobacter enzymogenes OH11 is a non-flagellated soil bacterium that utilizes type IV pilus (T4P)-driven twitching motility to prey upon nearby fungi for food. Interestingly, the strain OH11 encodes components homologous to the flagellar type III protein apparatus (FT3SS) on its genome, but it remains unknown whether this FT3SS-like system is functional. Here, we report that, despite the absence of flagella, the FT3SS homologous genes are responsible not only for the export of the heterologous flagellin in strain OH11 but also for twitching motility. Blocking the FT3SS-like system by in-frame deletion mutations in either flhB or fliI abolished the secretion of heterologous flagellin molecules into the culture medium, indicating that the FT3SS is functional in strain OH11. A deletion of flhA, flhB, fliI, or fliR inhibited T4P-driven twitching motility, whereas neither that of fliP nor fliQ did, suggesting that FlhA, FlhB, FliI, and FliR may obtain a novel function to modulate the twitching motility. The flagellar FliI ATPase was required for the secretion of the major pilus subunit, PilA, suggesting that FliI would have evolved to act as a PilB-like pilus ATPase. These observations lead to a plausible hypothesis that the non-flagellated L. enzymogenes OH11 could preserve FT3SS-like genes for acquiring a distinct function to regulate twitching motility associated with its predatory behavior.
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Thomas, Nikhil A., and Ken F. Jarrell. "Characterization of Flagellum Gene Families of Methanogenic Archaea and Localization of Novel Flagellum Accessory Proteins." Journal of Bacteriology 183, no. 24 (December 15, 2001): 7154–64. http://dx.doi.org/10.1128/jb.183.24.7154-7164.2001.
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ABSTRACT Archaeal flagella are unique motility structures, and the absence of bacterial structural motility genes in the complete genome sequences of flagellated archaeal species suggests that archaeal flagellar biogenesis is likely mediated by novel components. In this study, a conserved flagellar gene family from each of Methanococcus voltae, Methanococcus maripaludis,Methanococcus thermolithotrophicus, andMethanococcus jannaschii has been characterized. These species possess multiple flagellin genes followed immediately by eight known and supposed flagellar accessory genes,flaCDEFGHIJ. Sequence analyses identified a conserved Walker box A motif in the putative nucleotide binding proteins FlaH and FlaI that may be involved in energy production for flagellin secretion or assembly. Northern blotting studies demonstrated that all the species have abundant polycistronic mRNAs corresponding to some of the structural flagellin genes, and in some cases several flagellar accessory genes were shown to be cotranscribed with the flagellin genes. Cloned flagellar accessory genes of M. voltaewere successfully overexpressed as His-tagged proteins inEscherichia coli. These recombinant flagellar accessory proteins were affinity purified and used as antigens to raise polyclonal antibodies for localization studies. Immunoblotting of fractionated M. voltae cells demonstrated that FlaC, FlaD, FlaE, FlaH, and FlaI are all present in the cell as membrane-associated proteins but are not major components of isolated flagellar filaments. Interestingly, flaD was found to encode two proteins, each translated from a separate ribosome binding site. These protein expression data indicate for the first time that the putative flagellar accessory genes of M. voltae, and likely those of other archaeal species, do encode proteins that can be detected in the cell.