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Автор: Grafiati

Опубліковано: 6 вересня 2023

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1

Casali, Nicola, Amy M. White, and Lee W. Riley. "Regulation of the Mycobacterium tuberculosis mce1 Operon." Journal of Bacteriology 188, no. 2 (January 15, 2006): 441–49. http://dx.doi.org/10.1128/jb.188.2.441-449.2006.

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ABSTRACT In the murine model of infection, a Mycobacterium tuberculosis mce1 operon mutant elicits an aberrant granulomatous response, resulting in uncontrolled replication and failure to enter a persistent state. In this study, we demonstrate that the mce1 genes can be transcribed as a 13-gene polycistronic message encompassing Rv0166 to Rv0178. Quantitative reverse transcriptase PCR and immunoblot analyses revealed that the mce1 genes and proteins are expressed during in vitro growth but are significantly down-regulated in intracellular bacilli isolated from murine macrophages. A homologue of the FadR subfamily of GntR transcriptional regulators, Rv0165c (designated Mce1R), lies upstream and is divergently transcribed from the operon. To investigate whether this gene plays a role in regulation of mce1 expression, we created an M. tuberculosis mce1R deletion mutant. There was no difference in expression of mce1 operon genes in Δmce1R compared to expression in the wild type during logarithmic growth in vitro. However, in bacilli isolated from murine macrophages, expression of mce1 genes was significantly higher in Δmce1R. In addition, overexpression of mce1R resulted in repression of the mce1 genes. These data demonstrate that Mce1R is a negative regulator that acts intracellularly to repress expression of the mce1 operon. We propose that Mce1R facilitates balanced temporal expression of the mce1 products required for organized granuloma formation, which is both protective to the host and necessary for the persistence of M. tuberculosis.

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2

Sun, Y., Y. Wang, H. Wang, S. Wang, G. Ping, and L. Zhang. "HLA-A*0201-restricted CTL epitopes in Rv0350 and Rv0351 of latent Mycobacterium tuberculosis." International Journal of Infectious Diseases 21 (April 2014): 306. http://dx.doi.org/10.1016/j.ijid.2014.03.1054.

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3

Xu, Guangxian, Hao Jia, Yong Li, Xiaoming Liu, Min Li, and Yujiong Wang. "Hemolytic phospholipaseRv0183of Mycobacterium tuberculosis induces inflammatory response and apoptosis in alveolar macrophage RAW264.7 cells." Canadian Journal of Microbiology 56, no. 11 (November 2010): 916–24. http://dx.doi.org/10.1139/w10-079.

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The metabolic pathway of phospholipids is one of the most important physiologic pathways in Mycobacterium tuberculosis , a typical intracellular bacterium. The hemolytic phospholipase lip gene (Rv0183) is one of 24 phospholipase genes that have been demonstrated to play critical roles in the metabolism of phospholipids in M. tuberculosis. Quantitative RT–PCR and flow cytometry were used to elucidate the immunological and pathogenic implications of the Rv0183 gene on the inflammatory response following persistent expression of Rv0183 in mouse alveolar macrophage RAW264.7 cells. Our results demonstrate that a time-course-dependent ectopic expression of Rv0183 significantly elevated the expression of IL-6, NF-κB, TLR-2, TLR-6, TNFα, and MyD88 in these alveolar macrophage cells. Furthermore, the persistent expression of Rv0183 induced RAW264.7 cell apoptosis in vitro. These findings demonstrate that the expression of Rv0183 induces an inflammatory response and cell apoptosis in the host cells, suggesting that Rv0183 may play an important role in the virulence and pathogenesis of M. tuberculosis infection.

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4

Goldstone, Rachael M., Sunali D. Goonesekera, Barry R. Bloom, and Samantha L. Sampson. "The Transcriptional Regulator Rv0485 Modulates the Expression of a pe and ppe Gene Pair and Is Required for Mycobacterium tuberculosis Virulence." Infection and Immunity 77, no. 10 (August 3, 2009): 4654–67. http://dx.doi.org/10.1128/iai.01495-08.

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ABSTRACT The pe and ppe genes are unique to mycobacteria and are widely speculated to play a role in tuberculosis pathogenesis. However, little is known about how expression of these genes is controlled. Elucidating the regulatory control of genes found exclusively in mycobacteria, such as the pe and ppe gene families, may be key to understanding the success of this pathogen. In this study, we used a transposon mutagenesis approach to elucidate pe and ppe regulation. This resulted in the identification of Rv0485, a previously uncharacterized transcriptional regulator. Microarray and quantitative real-time PCR analysis confirmed that disruption of Rv0485 reduced the expression of the pe13 and ppe18 gene pair (Rv1195 and Rv1196), defined the Rv0485 regulon, and emphasized the lack of global regulation of pe and ppe genes. The in vivo phenotype of the Rv0485 transposon mutant strain (Rv0485::Tn) was investigated in the mouse model, where it was demonstrated that the mutation has minimal effect on bacterial organ burden. Despite this, disruption of Rv0485 allowed mice to survive for significantly longer, with substantially reduced lung pathology in comparison with mice infected with wild-type Mycobacterium tuberculosis. Infection of immune-deficient SCID mice with the Rv0485::Tn strain also resulted in extended survival times, suggesting that Rv0485 plays a role in modulation of innate immune responses. This is further supported by the finding that disruption of Rv0485 resulted in reduced secretion of proinflammatory cytokines by infected murine macrophages. In summary, we have demonstrated that disruption of a previously uncharacterized transcriptional regulator, Rv0485, results in reduced expression of pe13 and ppe18 and attenuation of M. tuberculosis virulence.

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5

Côtes, Karen, Rabeb Dhouib, Isabelle Douchet, Henri Chahinian, Alain de Caro, Frédéric Carrière, and Stéphane Canaan. "Characterization of an exported monoglyceride lipase from Mycobacterium tuberculosis possibly involved in the metabolism of host cell membrane lipids." Biochemical Journal 408, no. 3 (November 28, 2007): 417–27. http://dx.doi.org/10.1042/bj20070745.

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The Rv0183 gene of the Mycobacterium tuberculosis H37Rv strain, which has been implicated as a lysophospholipase, was cloned and expressed in Escherichia coli. The purified Rv0183 protein did not show any activity when lysophospholipid substrates were used, but preferentially hydrolysed monoacylglycerol substrates with a specific activity of 290 units·mg−1 at 37 °C. Rv0183 hydrolyses both long chain di- and triacylglycerols, as determined using the monomolecular film technique, although the turnover was lower than with MAG (monoacyl-glycerol). The enzyme shows an optimum activity at pH values ranging from 7.5 to 9.0 using mono-olein as substrate and is inactivated by serine esterase inhibitors such as E600, PMSF and tetrahydrolipstatin. The catalytic triad is composed of Ser110, Asp226 and His256 residues, as confirmed by the results of site-directed mutagenesis. Rv0183 shows 35% sequence identity with the human and mouse monoglyceride lipases and well below 15% with the other bacterial lipases characterized so far. Homologues of Rv0183 can be identified in other mycobacterial genomes such as Mycobacterium bovis, Mycobacterium smegmatis, and even Mycobacterium leprae, which is known to contain a low number of genes involved in the replication process within the host cells. The results of immunolocalization studies performed with polyclonal antibodies raised against the purified recombinant Rv0183 suggested that the enzyme was present only in the cell wall and culture medium of M. tuberculosis. Our results identify Rv0183 as the first exported lipolytic enzyme to be characterized in M. tuberculosis and suggest that Rv0183 may be involved in the degradation of the host cell lipids.

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6

Abomoelak, Bassam, Elizabeth A. Hoye, Jing Chi, Sarah A. Marcus, Francoise Laval, John P. Bannantine, Sarah K. Ward, Mamadou Daffé, Hong Di Liu, and Adel M. Talaat. "mosR, a Novel Transcriptional Regulator of Hypoxia and Virulence in Mycobacterium tuberculosis." Journal of Bacteriology 191, no. 19 (July 31, 2009): 5941–52. http://dx.doi.org/10.1128/jb.00778-09.

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ABSTRACT Latent tuberculosis represents a high-risk burden for one-third of the world population. Previous analysis of murine tuberculosis identified a novel transcriptional regulator encoded by Rv0348 that could control the establishment of persistent tuberculosis. Disruption of the Rv0348 gene from the genome of the virulent H37Rv strain of Mycobacterium tuberculosis revealed a global impact on the transcriptional profiles of 163 genes, including induction of the mammalian cell entry (mce1) operon and the repression of a significant number of genes involved in hypoxia and starvation responses. Nonetheless, gel shift assays did not reveal direct binding between Rv0348 and a set of regulated promoters, suggesting an indirect regulatory role. However, when expressed in Mycobacterium smegmatis, the Rv0348 transcripts were significantly responsive to different levels of hypoxia and the encoded protein was shown to regulate genes involved in hypoxia [e.g., Rv3130c (tgs1)] and intracellular survival (e.g., mce1), among other genes. Interestingly, the colonization level of the ΔmosR mutant strain was significantly lower than that of the wild-type strain of M. tuberculosis, suggesting its attenuation in the murine model of tuberculosis. Taken together, our analyses indicated that the Rv0348 gene encodes a novel transcriptional factor that regulates several operons involved in mycobacterial survival, especially during hypoxia; hence, we propose that Rv0348 be renamed mosR for regulator of mycobacterial operons of survival.

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7

Sharma, Divakar, Manju Lata, Mohammad Faheem, Asad Khan, Beenu Joshi, Krishnamurthy Venkatesan, Sangeeta Shukla, and Deepa Bisht. "Cloning, Expression and Correlation of Rv0148 to Amikacin & Kanamycin Resistance." Current Proteomics 12, no. 2 (September 3, 2015): 96–100. http://dx.doi.org/10.2174/157016461202150903113053.

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8

Gurvitz, Aner, J. Kalervo Hiltunen, and Alexander J. Kastaniotis. "Heterologous Expression of Mycobacterial Proteins in Saccharomyces cerevisiae Reveals Two Physiologically Functional 3-Hydroxyacyl-Thioester Dehydratases, HtdX and HtdY, in Addition to HadABC and HtdZ." Journal of Bacteriology 191, no. 8 (January 9, 2009): 2683–90. http://dx.doi.org/10.1128/jb.01046-08.

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ABSTRACT We report on Mycobacterium tuberculosis Rv0241c and Rv3389c, representing two physiologically functional 3-hydroxyacyl-thioester dehydratases (Htd). These enzymes are potentially entrained in type 2 fatty acid synthase (FASII). Mycobacterial FASII is involved in the synthesis of mycolic acids, which are the major constituents of the protective layer around the pathogen, shielding it from noxious chemicals and the host's immune system. Mycolic acids are additionally associated with the virulence and resilience of M. tuberculosis. Here, Rv0241c and Rv3389c, which are distinct from the previously identified heterodimers Rv0635-Rv0636 (HadAB) and Rv0636-Rv0637 (HadBC) but also the homodimer Rv0130 (HtdZ), were identified by expressing the corresponding candidate open reading frames in Saccharomyces cerevisiae htd2Δ cells lacking mitochondrial 3-hydroxyacyl-acyl carrier protein dehydratase activity, followed by scoring for phenotype rescue. The htd2Δ mutant fails to produce sufficient levels of lipoic acid and does not respire or grow on nonfermentable carbon sources. Soluble protein extracts made from mutant htd2Δ cells expressing mitochondrially targeted Rv0241c or Rv3389c contained 3-hydroxyacyl-thioester hydratase activity. Moreover, mutant yeast cells expressing Rv0241c or Rv3389c were able to recover their respiratory growth on glycerol medium and efficiently reduce 2,3,5-triphenyltetrazolium chloride. Additionally, expression of mitochondrial Rv0241c or Rv3389c in htd2Δ cells also restored de novo lipoic acid synthesis to 92 and 40% of the level in the wild-type strain, respectively. We propose naming Rv0241c and Rv3389c as HtdX and HtdY, respectively, and discuss the implications of our finding with reference to Rv0098, a candidate mycobacterial FabZ homologue with intrinsic thioesterase and hydratase activities that lacks the eukaryotic-like hydratase-2 motif.

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9

Anlay, Degefaye Zelalem, Emmanuel Rivière, Pham Hien Trang Tu, Steven Abrams, and Annelies Van Rie. "A Bayesian approach to estimate the probability of resistance to bedaquiline in the presence of a genomic variant." PLOS ONE 18, no. 6 (June 14, 2023): e0287019. http://dx.doi.org/10.1371/journal.pone.0287019.

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Background Bedaquiline is a core drug for treatment of rifampicin-resistant tuberculosis. Few genomic variants have been statistically associated with bedaquiline resistance. Alternative approaches for determining the genotypic-phenotypic association are needed to guide clinical care. Methods Using published phenotype data for variants in Rv0678, atpE, pepQ and Rv1979c genes in 756 Mycobacterium tuberculosis isolates and survey data of the opinion of 33 experts, we applied Bayesian methods to estimate the posterior probability of bedaquiline resistance and corresponding 95% credible intervals. Results Experts agreed on the role of Rv0678, and atpE, were uncertain about the role of pepQ and Rv1979c variants and overestimated the probability of bedaquiline resistance for most variant types, resulting in lower posterior probabilities compared to prior estimates. The posterior median probability of bedaquiline resistance was low for synonymous mutations in atpE (0.1%) and Rv0678 (3.3%), high for missense mutations in atpE (60.8%) and nonsense mutations in Rv0678 (55.1%), relatively low for missense (31.5%) mutations and frameshift (30.0%) in Rv0678 and low for missense mutations in pepQ (2.6%) and Rv1979c (2.9%), but 95% credible intervals were wide. Conclusions Bayesian probability estimates of bedaquiline resistance given the presence of a specific mutation could be useful for clinical decision-making as it presents interpretable probabilities compared to standard odds ratios. For a newly emerging variant, the probability of resistance for the variant type and gene can still be used to guide clinical decision-making. Future studies should investigate the feasibility of using Bayesian probabilities for bedaquiline resistance in clinical practice.

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10

Grininger, Christoph, Mario Leypold, Philipp Aschauer, Tea Pavkov-Keller, Lina Riegler-Berket, Rolf Breinbauer, and Monika Oberer. "Structural Changes in the Cap of Rv0183/mtbMGL Modulate the Shape of the Binding Pocket." Biomolecules 11, no. 9 (September 1, 2021): 1299. http://dx.doi.org/10.3390/biom11091299.

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Tuberculosis continues to be a major threat to the human population. Global efforts to eradicate the disease are ongoing but are hampered by the increasing occurrence of multidrug-resistant strains of Mycobacterium tuberculosis. Therefore, the development of new treatment, and the exploration of new druggable targets and treatment strategies, are of high importance. Rv0183/mtbMGL, is a monoacylglycerol lipase of M. tuberculosis and it is involved in providing fatty acids and glycerol as building blocks and as an energy source. Since the lipase is expressed during the dormant and active phase of an infection, Rv0183/mtbMGL is an interesting target for inhibition. In this work, we determined the crystal structures of a surface-entropy reduced variant K74A Rv0183/mtbMGL in its free form and in complex with a substrate mimicking inhibitor. The two structures reveal conformational changes in the cap region that forms a major part of the substrate/inhibitor binding region. We present a completely closed conformation in the free form and semi-closed conformation in the ligand-bound form. These conformations differ from the previously published, completely open conformation of Rv0183/mtbMGL. Thus, this work demonstrates the high conformational plasticity of the cap from open to closed conformations and provides useful insights into changes in the substrate-binding pocket, the target of potential small-molecule inhibitors.

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11

Okkels, Limei Meng, and Peter Andersen. "Protein-Protein Interactions of Proteins from the ESAT-6 Family of Mycobacterium tuberculosis." Journal of Bacteriology 186, no. 8 (April 15, 2004): 2487–91. http://dx.doi.org/10.1128/jb.186.8.2487-2491.2004.

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ABSTRACT In the present study, we demonstrate that, in analogy with the genes encoding ESAT-6 and CFP-10, the genes rv0287 and rv0288 from the ESAT-6 gene family are cotranscribed. Using Western-Western blotting and protein-print overlay methodologies, we demonstrate that ESAT-6 and CFP-10, as well as the protein pair Rv0288/Rv0287, interact pairwise in a highly specific way. Most notably, the ESAT-6 proteins interact directly with Rv3873, a possible cell envelope component of the ESAT-6 secretion pathway.

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12

Bondoc, Jasper Marc G., Hiten J. Gutka, Mashal M. Almutairi, Ryan Patwell, Maxwell W. Rutter, Nina M. Wolf, Ram Samudrala, Shahila Mehboob, and Farahnaz Movahedzadeh. "Rv0100, a proposed acyl carrier protein in Mycobacterium tuberculosis: expression, purification and crystallization." Acta Crystallographica Section F Structural Biology Communications 75, no. 10 (September 24, 2019): 646–51. http://dx.doi.org/10.1107/s2053230x19012652.

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Acyl carrier proteins (ACPs) are important components in fatty-acid biosynthesis in prokaryotes. Rv0100 is predicted to be an essential ACP in Mycobacterium tuberculosis, the pathogen that is the causative agent of tuberculosis, and therefore has the potential to be a novel antituberculosis drug target. Here, the successful cloning and purification of Rv0100 using Mycobacterium smegmatis as a host is reported. Crystals of the purified protein were obtained that diffracted to a resolution of 1.9 Å. Overall, this work lays the foundation for the future pursuit of drug discovery and development against this potentially novel drug target.

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13

Bhargavi, Gunapati, Amit Kumar Singh, Shripad A. Patil, and Kannan Palaniyandi. "A putative short-chain dehydrogenase Rv0148 of Mycobacterium tuberculosis affects bacterial survival and virulence." Current Research in Microbial Sciences 3 (2022): 100113. http://dx.doi.org/10.1016/j.crmicr.2022.100113.

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14

Dhouib, Rabeb, Françoise Laval, Frédéric Carrière, Mamadou Daffé, and Stéphane Canaan. "A Monoacylglycerol Lipase from Mycobacterium smegmatis Involved in Bacterial Cell Interaction." Journal of Bacteriology 192, no. 18 (July 2, 2010): 4776–85. http://dx.doi.org/10.1128/jb.00261-10.

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ABSTRACT MSMEG_0220 from Mycobacterium smegmatis, the ortholog of the Rv0183 gene from M. tuberculosis, recently identified and characterized as encoding a monoacylglycerol lipase, was cloned and expressed in Escherichia coli. The recombinant protein (rMSMEG_0220), which exhibits 68% amino acid sequence identity with Rv0183, showed the same substrate specificity and similar patterns of pH-dependent activity and stability as the M. tuberculosis enzyme. rMSMEG_0220 was found to hydrolyze long-chain monoacylglycerol with a specific activity of 143 ± 6 U mg−1. Like Rv0183 in M. tuberculosis, MSMEG_0220 was found to be located in the cell wall. To assess the in vivo role of the homologous proteins, an MSMEG_0220 disrupted mutant of M. smegmatis (MsΔ0220) was produced. An intriguing change in the colony morphology and in the cell interaction, which were partly restored in the complemented mutant containing either an active (ComMsΔ0220) or an inactive (ComMsΔ0220S111A) enzyme, was observed. Growth studies performed in media supplemented with monoolein showed that the ability of both MsΔ0220 and ComMsΔ0220S111A to grow in the presence of this lipid was impaired. Moreover, studies of the antimicrobial susceptibility of the MsΔ0220 strain showed that this mutant is more sensitive to rifampin and more resistant to isoniazid than the wild-type strain, pointing to a critical structural role of this enzyme in mycobacterial physiology, in addition to its function in the hydrolysis of exogenous lipids.

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15

Omar, Shaheed V., Farzana Ismail, Norbert Ndjeka, Koné Kaniga, and Nazir A. Ismail. "Bedaquiline-Resistant Tuberculosis Associated with Rv0678 Mutations." New England Journal of Medicine 386, no. 1 (January 6, 2022): 93–94. http://dx.doi.org/10.1056/nejmc2103049.

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16

Radhakrishnan, Abhijith, Nitin Kumar, Catherine C. Wright, Tsung-Han Chou, Marios L. Tringides, Jani Reddy Bolla, Hsiang-Ting Lei, et al. "Crystal Structure of the Transcriptional Regulator Rv0678 ofMycobacterium tuberculosis." Journal of Biological Chemistry 289, no. 23 (April 15, 2014): 16526–40. http://dx.doi.org/10.1074/jbc.m113.538959.

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17

Danilchanka, Olga, Claudia Mailaender, and Michael Niederweis. "Identification of a Novel Multidrug Efflux Pump of Mycobacterium tuberculosis." Antimicrobial Agents and Chemotherapy 52, no. 7 (May 5, 2008): 2503–11. http://dx.doi.org/10.1128/aac.00298-08.

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ABSTRACT The impermeability of the outer membrane in combination with drug efflux are major determinants of the natural drug resistance of mycobacteria. β-Lactams are the most widely used antibiotics for treatment of bacterial infections. However, it is unknown how β-lactams enter Mycobacterium tuberculosis and whether efflux pumps exist that can export these drugs out of the cell. To identify the molecular mechanisms of M. tuberculosis resistance to β-lactams, a library of 7,500 transposon mutants was generated in the model organism Mycobacterium bovis BCG. Thirty-three unique insertion sites were determined that conferred medium or high-level (≥2,000 μg/ml) resistance to ampicillin. Three mutants in sulfolipid synthesis or transport were highly resistant to ampicillin, indicating an indirect effect of the lipid composition on the outer membrane permeability of M. bovis BCG to ampicillin. Mutants with insertions in genes encoding surface molecules such as PPE proteins or lipoarabinomannan were also completely resistant to ampicillin, thus suggesting a lack of transport across the outer membrane. Insertion of the transposon in front of bcg0231 increased transcription of the gene and concomitantly the resistance of M. bovis BCG to ampicillin, streptomycin, and chloramphenicol by 32- to 64-fold. Resistance to vancomycin and tetracycline was increased four- to eightfold. Bcg0231 and Rv0194 are almost identical ATP-binding cassette transporters. Expression of rv0194 significantly reduced accumulation of ethidium bromide and conferred multidrug resistance to Mycobacterium smegmatis. Both effects were abrogated in the presence of the efflux pump inhibitor reserpine. These results demonstrate that Rv0194 is a novel multidrug efflux pump of M. tuberculosis.

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18

Park, Sanghee, Jihee Jung, Jiyeon Kim, Sang Bong Han, and Sungweon Ryoo. "Investigation of Clofazimine Resistance and Genetic Mutations in Drug-Resistant Mycobacterium tuberculosis Isolates." Journal of Clinical Medicine 11, no. 7 (March 30, 2022): 1927. http://dx.doi.org/10.3390/jcm11071927.

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Recently, as clofazimine (CFZ) showed a good therapeutic effect in treating multi-drug-resistant tuberculosis (MDR-TB), the anti-tuberculosis activity and resistance were re-focused. Here, we investigated the CFZ resistance and genetic mutations of drug-resistant Mycobacterium tuberculosis (DR-Mtb) isolates to improve the diagnosis and treatment of drug-resistant TB patients. The minimal inhibitory concentration (MIC) of CFZ was examined by resazurin microtiter assay (REMA) with two reference strains and 122 clinical isolates from Korea. The cause of CFZ resistance was investigated in relation to the therapeutic history of patients. Mutations of Rv0678, Rv1979c and pepQ of CFZ resistant isolates were analyzed by PCR and DNA sequencing. The rate of CFZ resistance with MIC > 1 mg/L was 4.1% in drug-resistant Mtb isolates. The cause of CFZ resistance was not related to treatment with CFZ or bedaquiline. A CFZ susceptibility test should be conducted regardless of dugs use history. The four novel mutation sites were identified in the Rv0678 and pepQ genes related to CFZ resistance in this study.

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19

Meikle, V., A. Alito, A. S. Llera, A. Gioffré, A. Peralta, B. M. Buddle, and A. Cataldi. "Identification of Novel Mycobacterium bovis Antigens by Dissection of Crude Protein Fractions." Clinical and Vaccine Immunology 16, no. 9 (July 29, 2009): 1352–59. http://dx.doi.org/10.1128/cvi.00211-09.

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ABSTRACT Culture filtrate and cell extracts from Mycobacterium bovis cultures contain molecules which could promote protective immunity to tuberculosis in animals. Different protein fractions of M. bovis cultures were obtained by elution electrophoresis and were tested in experimentally infected cattle. The fractions that elicited gamma interferon (IFN-γ) responses were resolved by two-dimensional gel electrophoresis, and individual proteins were identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry. The open reading frames were cloned, expressed as their recombinant forms, and retested with naturally and experimentally infected animals. Eleven protein fractions were highly reactive, from which the Rv1636, HspX, Rv0138, Rv2524, EsxI, and Rv3740 recombinant proteins were obtained. EsxI and HspX were the antigens most recognized by the IFN-γ release assay. In summary, a proteomic approach allowed the identification of novel antigens useful for the diagnosis of bovine tuberculosis.

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20

Mishra, Arun K., Sarah Batt, Karin Krumbach, Lothar Eggeling та Gurdyal S. Besra. "Characterization of the Corynebacterium glutamicum ΔpimB′ ΔmgtA Double Deletion Mutant and the Role of Mycobacterium tuberculosis Orthologues Rv2188c and Rv0557 in Glycolipid Biosynthesis". Journal of Bacteriology 191, № 13 (24 квітня 2009): 4465–72. http://dx.doi.org/10.1128/jb.01729-08.

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ABSTRACT In this study, utilizing a Corynebacterium glutamicum ΔpimB′ ΔmgtA double deletion mutant, we unequivocally assign the in vivo functions of Rv2188c as an Ac1PIM1:mannosyltransferase (originally termed PimB′ Mt [Mycobacterium tuberculosis PimB′]) and Rv0557 as a GlcAGroAc2:mannosyltransferase (originally termed PimB Mt ), which we have reassigned as PimB Mt and MgtA Mt , respectively, in Mycobacterium tuberculosis.

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21

Johansson, Patrik, Alina Castell, T. Alwyn Jones, and Kristina Bäckbro. "Structure and function of Rv0130, a conserved hypothetical protein fromMycobacterium tuberculosis." Protein Science 15, no. 10 (October 2006): 2300–2309. http://dx.doi.org/10.1110/ps.062309306.

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22

Muttucumaru, D. G. Niranjala, Debbie A. Smith, Elizabeth J. McMinn, Valerie Reese, Rhea N. Coler, and Tanya Parish. "Mycobacterium tuberculosis Rv0198c, a putative matrix metalloprotease is involved in pathogenicity." Tuberculosis 91, no. 2 (March 2011): 111–16. http://dx.doi.org/10.1016/j.tube.2010.11.010.

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23

Fang, Haihong, Dan Yu, Yuzhi Hong, Xiaodan Zhou, Chuanyou Li, and Baolin Sun. "The LuxR family regulator Rv0195 modulates Mycobacterium tuberculosis dormancy and virulence." Tuberculosis 93, no. 4 (July 2013): 425–31. http://dx.doi.org/10.1016/j.tube.2013.04.005.

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24

Gurvitz, Aner, J. Kalervo Hiltunen, and Alexander J. Kastaniotis. "Identification of a Novel Mycobacterial 3-Hydroxyacyl-Thioester Dehydratase, HtdZ (Rv0130), by Functional Complementation in Yeast." Journal of Bacteriology 190, no. 11 (March 28, 2008): 4088–90. http://dx.doi.org/10.1128/jb.00016-08.

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ABSTRACT We report on the identification of Mycobacterium tuberculosis HtdZ (Rv0130), representing a novel 3-hydroxyacyl-thioester dehydratase. HtdZ was picked up by the functional complementation of Saccharomyces cerevisiae htd2Δ cells lacking the dehydratase of mitochondrial type II fatty acid synthase. Mutant cells expressing HtdZ contained dehydratase activity, recovered their respiratory ability, and partially restored de novo lipoic acid synthesis.

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Alexander, David, Joshua Yoneda, Supriya Bhat, Teagan Oleksyn, Jeffrey Chen, and Cameron Andrew. "1828. Bedaquiline Resistance in Mycobacterium intracellulare Is Mediated by the Transcriptional Repressor MmpT5." Open Forum Infectious Diseases 6, Supplement_2 (October 2019): S41. http://dx.doi.org/10.1093/ofid/ofz359.090.

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Abstract Background Bedaquiline (BDQ) is an FDA approved antibiotic with antimycobacterial activity. BDQ resistance has been observed in several Mycobacterium species. High-level resistance is due to mutations in ATP synthase. Low -level resistance is attributed to drug efflux. Previously, we suggested that the MmpSL5 efflux system mediates BDQ resistance in M. intracellulare. Here, we examine the role of MmpT5 in transcriptional regulation of mmpSL5 and BDQ resistance. Methods In this study, mmpSL5-mmpT5 genes were cloned from 2 pre-treatment (wild-type mmpT5) and 2 relapse (mutant mmpT5) isolates of M. intracellulare and transformed into M. smegmatis. BDQ MICs were determined as well as cell survival after 24 hours exposure to an inhibitory concentration (0.07 µg/mL) of BDQ. Transcription of the M. intracellulare mmpT5 and mmpSL5 promoters was monitored with luciferase reporter gene fusions in the presence of wild-type and mutant alleles of mmpT5. Single and multigene constructs were created using the MoClo system, and transformed into E. coli DH5α. Constructs containing the M. tuberculosis rv0678 gene, which mediates low-level BDQ resistance in M. tuberculosis, were also examined. Results The BDQ MIC for the M. smegmatis control strain, and all strains containing mmpSL5-mmpT5 constructs, was 0.007 µg/mL. Even so, strains containing mutant mmpT5 alleles showed enhanced survival after 24 hours exposure to 0.007 µg/mL BDQ. Bacterial colonies associated with mutant mmpT5 alleles exhibited altered morphology relative to wild-type strains. Transcription of mmpSL5 was repressed by wild-type mmpT5, but neither mutant mmpT5 nor rv0678 repressed transcription. The mmpT5 luciferase reporter was not active. Conclusion MmpT5 represses transcription of mmpSL5 whereas the operon is dysregulated by mmpT5 mutations. Although Rv0678 regulates mmpSL expression in M. tuberculosis, it cannot repress the M. intracellulare mmpSL5 genes. The mmpSL5-mmpT5 genes have no impact on the BDQ MIC for M. smegmatis, but constructs containing mutant mmpT5 alleles do enhance bacterial survival. The altered morphology of these colonies suggests that BDQ resistance is mediated by cell wall changes in combination with drug efflux. Disclosures All Authors: No reported Disclosures.

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26

Sharma, Divakar, Rananjay Singh, Nirmala Deo, and Deepa Bisht. "Interactome analysis of Rv0148 to predict potential targets and their pathways linked to aminoglycosides drug resistance: An insilico approach." Microbial Pathogenesis 121 (August 2018): 179–83. http://dx.doi.org/10.1016/j.micpath.2018.05.034.

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Kaur, Jashandeep, and Jagdeep Kaur. "Rv0518, a nutritive stress inducible GDSL lipase of Mycobacterium tuberculosis, enhanced intracellular survival of bacteria by cell wall modulation." International Journal of Biological Macromolecules 135 (August 2019): 180–95. http://dx.doi.org/10.1016/j.ijbiomac.2019.05.121.

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Saravanan, Parameswaran, Vikash Kumar Dubey, and Sanjukta Patra. "Potential Selective Inhibitors against Rv0183 of Mycobacterium tuberculosis Targeting Host Lipid Metabolism." Chemical Biology & Drug Design 79, no. 6 (April 17, 2012): 1056–62. http://dx.doi.org/10.1111/j.1747-0285.2012.01373.x.

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Abrahams, Katherine A., Wei Hu, Gang Li, Yu Lu, Emily J. Richardson, Nicholas J. Loman, Haihong Huang, and Gurdyal S. Besra. "Anti-tubercular derivatives of rhein require activation by the monoglyceride lipase Rv0183." Cell Surface 6 (December 2020): 100040. http://dx.doi.org/10.1016/j.tcsw.2020.100040.

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Jones, Gareth J., Chris Pirson, Hannah P. Gideon, Katalin A. Wilkinson, David R. Sherman, Robert J. Wilkinson, R. Glyn Hewinson, and H. Martin Vordermeier. "Immune Responses to the Enduring Hypoxic Response Antigen Rv0188 Are Preferentially Detected in Mycobacterium bovis Infected Cattle with Low Pathology." PLoS ONE 6, no. 6 (June 21, 2011): e21371. http://dx.doi.org/10.1371/journal.pone.0021371.

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Yousuf, Suhail, Rajendra Kumar Angara, Ajit Roy, Shailesh Kumar Gupta, Rohan Misra, and Akash Ranjan. "Mce2R/Rv0586 of Mycobacterium tuberculosis is the functional homologue of FadR E. coli." Microbiology 164, no. 9 (September 1, 2018): 1133–45. http://dx.doi.org/10.1099/mic.0.000686.

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Saravanan, Parameswaran, and Sanjukta Patra. "Discovery of Potential Dual Inhibitors Against Lipases Rv0183 and Rv3802c for Tuberculosis Therapeutics." Letters in Drug Design & Discovery 13, no. 2 (November 12, 2015): 185–95. http://dx.doi.org/10.2174/1570180812999150812165215.

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Ouni, Rym, Houda Gharsalli, Violette Dirix, Amani Braiek, Nadia Sendi, Afifa Jarraya, Leila Douik El Gharbi, Mohamed‐Ridha Barbouche, and Chaouki Benabdessalem. "Granzyme B induced by Rv0140 antigen discriminates latently infected from active tuberculosis individuals." Journal of Leukocyte Biology 105, no. 2 (September 13, 2018): 297–306. http://dx.doi.org/10.1002/jlb.ma0318-117r.

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34

Bondoc, Jasper Marc G., Hiten J. Gutka, Mashal M. Almutairi, Ryan Patwell, Maxwell W. Rutter, Nina M. Wolf, Ram Samudrala, et al. "Rv0100, a proposed acyl carrier protein in Mycobacterium tuberculosis: expression, purification and crystallization. Corrigendum." Acta Crystallographica Section F Structural Biology Communications 76, no. 4 (April 1, 2020): 192–93. http://dx.doi.org/10.1107/s2053230x2000271x.

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35

Kaniga, K., N. Lounis, S. Zhuo, N. Bakare, and K. Andries. "Impact of Rv0678 mutations on patients with drug-resistant TB treated with bedaquiline." International Journal of Tuberculosis and Lung Disease 26, no. 6 (June 1, 2022): 571–73. http://dx.doi.org/10.5588/ijtld.21.0670.

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36

Weyand, Simone, Georgia Kefala, and Manfred S. Weiss. "Cloning, expression, purification, crystallization and preliminary X-ray diffraction analysis of DapC (Rv0858c) fromMycobacterium tuberculosis." Acta Crystallographica Section F Structural Biology and Crystallization Communications 62, no. 8 (July 25, 2006): 794–97. http://dx.doi.org/10.1107/s1744309106026753.

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37

Speer, Alexander, Jim Sun, Olga Danilchanka, Virginia Meikle, Jennifer L. Rowland, Kerstin Walter, Bradford R. Buck, et al. "Surface hydrolysis of sphingomyelin by the outer membrane protein Rv0888 supports replication ofMycobacterium tuberculosisin macrophages." Molecular Microbiology 97, no. 5 (July 4, 2015): 881–97. http://dx.doi.org/10.1111/mmi.13073.

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Johnston, Jodie M., Ming Jiang, Zhihong Guo, and Edward N. Baker. "Structural and functional analysis of Rv0554 fromMycobacterium tuberculosis: testing a putative role in menaquinone biosynthesis." Acta Crystallographica Section D Biological Crystallography 66, no. 8 (July 10, 2010): 909–17. http://dx.doi.org/10.1107/s0907444910025771.

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Mycobacterium tuberculosis, the cause of tuberculosis, is a devastating human pathogen against which new drugs are urgently needed. Enzymes from the biosynthetic pathway for menaquinone are considered to be valid drug targets. The protein encoded by the open reading frameRv0554has been expressed, purified and subjected to structural and functional analysis to test for a putative role in menaquinone biosynthesis. The crystal structure of Rv0554 has been solved and refined in two different space groups at 2.35 and 1.9 Å resolution. The protein is dimeric, with an α/β-hydrolase monomer fold. In each monomer, a large cavity adjacent to the catalytic triad is enclosed by a helical lid. Dimerization is mediated by the lid regions. Small-molecule additives used in crystallization bind in the active site, but no binding of ligands related to menaquinone biosynthesis could be detected and functional assays failed to support possible roles in menaquinone biosynthesis.

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39

Lee, Seung Jun, Sung Jae Shin, Moon Hee Lee, Min-Goo Lee, Tae Heung Kang, Won Sun Park, Byoung Yul Soh, et al. "A Potential Protein Adjuvant Derived from Mycobacterium tuberculosis Rv0652 Enhances Dendritic Cells-Based Tumor Immunotherapy." PLoS ONE 9, no. 8 (August 7, 2014): e104351. http://dx.doi.org/10.1371/journal.pone.0104351.

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Li, Xue, Ping Li, Cao Ruan, Long xiang Xie, Yinzhong Gu, Jiang Li, Qin Yi, Xi Lv, and Jianping Xie. "Mycobacterium tuberculosis Rv0191 is an efflux pump of major facilitator superfamily transporter regulated by Rv1353c." Archives of Biochemistry and Biophysics 667 (May 2019): 59–66. http://dx.doi.org/10.1016/j.abb.2019.04.010.

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41

Byun, Eui-Hong, Woo Sik Kim, A.-Rum Shin, Jong-Seok Kim, Jake Whang, Choul-Jae Won, Yohan Choi, et al. "Rv0315, a novel immunostimulatory antigen of Mycobacterium tuberculosis, activates dendritic cells and drives Th1 immune responses." Journal of Molecular Medicine 90, no. 3 (October 13, 2011): 285–98. http://dx.doi.org/10.1007/s00109-011-0819-2.

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42

Yang, Hongliang, JoLynn Troudt, Ajay Grover, Kimberly Arnett, Megan Lucas, Yun Sang Cho, Helle Bielefeldt-Ohmann, Jennifer Taylor, Angelo Izzo, and Karen M. Dobos. "Three Protein Cocktails Mediate Delayed-Type Hypersensitivity Responses Indistinguishable from That Elicited by Purified Protein Derivative in the Guinea Pig Model ofMycobacterium tuberculosisInfection." Infection and Immunity 79, no. 2 (December 6, 2010): 716–23. http://dx.doi.org/10.1128/iai.00486-10.

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ABSTRACTPurified protein derivative (PPD) is a widely used reagent for the diagnosis ofMycobacterium tuberculosisinfection. Recently, the molecular composition of PPD was defined, with hundreds of mycobacterial protein representatives making up PPD. Which, if any, of these specific products drive the potency of PPD remains in question. In this study, two proteins (DnaK and GroEL2) previously identified as dominant proteins in PPD were tested for the capacity to induce delayed-type hypersensitivity (DTH) responses in H37Rv-infected or BCG-vaccinated guinea pigs. These two proteins were used in pull-down assays to identify interacting PPD products. Six proteins were identified as interacting partners with DnaK and GroEL2, i.e., Rv0009, Rv0475, Rv0569, Rv0685, Rv2626c, and Rv2632c. These six proteins were tested alone and in combination with DnaK and GroEL2 for the capacity to induce a DTH response in the guinea pig model. From these studies, two cocktails, DnaK/GroEL2/Rv0009 and DnaK/GroEL2/Rv0685, were found to induce DTH responses in H37Rv-infected or BCG-vaccinated guinea pigs that were indistinguishable from DTH responses driven by a PPD injection. The mechanism by which DTH responses were induced was elucidated by histologic examination, analysis of activated CD4+/CD8+T cells, and cytokine mRNA expression at the site of the DTH response. PPD and the protein cocktails tested induced strong DTH responses in H37Rv-infected guinea pigs. Ex vivo phenotyping of T cells at the DTH site indicated that this response is mediated by activated CD4+and CD8+T cells, with increases in gamma interferon and tumor necrosis factor alpha, but not interleukin-10, at the site of the DTH response. Our results demonstrate for the first time that the PPD response can be mimicked at the molecular level with defined protein cocktails. The use of this defined product will allow a more thorough understanding of the DTH response and may provide a platform for more rapid and sensitive second-generation skin test reagents for the diagnosis ofM. tuberculosisinfection.

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43

Tiwari, Bhavana Mishra, Nisha Kannan, Lakshmi Vemu, and Tirumalai R. Raghunand. "The Mycobacterium tuberculosis PE Proteins Rv0285 and Rv1386 Modulate Innate Immunity and Mediate Bacillary Survival in Macrophages." PLoS ONE 7, no. 12 (December 17, 2012): e51686. http://dx.doi.org/10.1371/journal.pone.0051686.

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44

Liu, Yongliang, Xiaofei Li, Wei Liu, Yang Liu, Zhouyue Zhong, Lili Wang, ShengXiang Ge, Jun Zhang, and Ningshao Xia. "IL-6 release of Rv0183 antigen-stimulated whole blood is a potential biomarker for active tuberculosis patients." Journal of Infection 76, no. 4 (April 2018): 376–82. http://dx.doi.org/10.1016/j.jinf.2017.11.004.

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45

Cockle, P. J., S. V. Gordon, R. G. Hewinson, and H. M. Vordermeier. "Field Evaluation of a Novel Differential Diagnostic Reagent for Detection of Mycobacterium bovis in Cattle." Clinical and Vaccine Immunology 13, no. 10 (August 30, 2006): 1119–24. http://dx.doi.org/10.1128/cvi.00209-06.

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ABSTRACT In the search for improved tools with which to control bovine tuberculosis, the development of enhanced immunodiagnostic reagents is a high priority. Such reagents are required to improve the performance of tuberculin-based reagents and allow the discrimination of vaccinated cattle from those infected with Mycobacterium bovis. In this study, we identified the immunodominant, frequently recognized peptides from Rv3873, Rv3879c, Rv0288, and Rv3019c, which, together with peptides comprising the current lead diagnostic antigens, ESAT-6 and CFP-10, were formulated into a peptide cocktail. In a test of naturally infected cattle, this cocktail was significantly better than tuberculin was for identifying skin test-negative animals with confirmed bovine tuberculosis. In addition, the specificity of this cocktail was not compromised by Mycobacterium bovis BCG vaccination. In summary, our results prioritize this peptide-based, fully synthetic reagent for assessment in larger trials.

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46

Gao, Beilan, Jie Wang, Jing Huang, Xiaochen Huang, Wei Sha, and Lianhua Qin. "The dynamic region of the peptidoglycan synthase gene, Rv0050, induces the growth rate and morphologic heterogeneity in Mycobacteria." Infection, Genetics and Evolution 72 (August 2019): 86–92. http://dx.doi.org/10.1016/j.meegid.2018.12.012.

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47

Maity, Koustav, Preeti Bajaj, Namita Surolia, Avadhesha Surolia, and Kaza Suguna. "Insights into the Substrate Specificity of a Thioesterase Rv0098 ofMycobacterium Tuberculosisthrough X-ray Crystallographic and Molecular Dynamics Studies." Journal of Biomolecular Structure and Dynamics 29, no. 5 (April 2012): 973–83. http://dx.doi.org/10.1080/07391102.2012.10507417.

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48

Ilghari, Dariush, Lorna C. Waters, Vaclav Veverka, Frederick W. Muskett, and Mark D. Carr. "15N, 13C and 1H resonance assignments and secondary structure determination of the Mycobacterium tuberculosis Rv0287–Rv0288 protein complex." Biomolecular NMR Assignments 3, no. 2 (June 21, 2009): 171–74. http://dx.doi.org/10.1007/s12104-009-9167-3.

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49

Scherzinger, Daniel, Erdmann Scheffer, Cornelia Bär, Hansgeorg Ernst, and Salim Al-Babili. "The Mycobacterium tuberculosis ORF Rv0654 encodes a carotenoid oxygenase mediating central and excentric cleavage of conventional and aromatic carotenoids." FEBS Journal 277, no. 22 (October 4, 2010): 4662–73. http://dx.doi.org/10.1111/j.1742-4658.2010.07873.x.

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50

Shee, Somnath, Reshma T. Veetil, Karthikeyan Mohanraj, Mayashree Das, Nitish Malhotra, Devleena Bandopadhyay, Hussain Beig, et al. "Biosensor-integrated transposon mutagenesis reveals rv0158 as a coordinator of redox homeostasis in Mycobacterium tuberculosis." eLife 12 (August 29, 2023). http://dx.doi.org/10.7554/elife.80218.

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Mycobacterium tuberculosis (Mtb) is evolutionarily equipped to resist exogenous reactive oxygen species but shows vulnerability to an increase in endogenous ROS (eROS). Since eROS is an unavoidable consequence of aerobic metabolism, understanding how Mtb manages eROS levels is essential yet needs to be characterized. By combining the Mrx1-roGFP2 redox biosensor with transposon mutagenesis, we identified 368 genes (redoxosome) responsible for maintaining homeostatic levels of eROS in Mtb. Integrating redoxosome with a global network of transcriptional regulators revealed a hypothetical protein (Rv0158) as a critical node managing eROS in Mtb. Disruption of rv0158 (rv0158 KO) impaired growth, redox balance, respiration, and metabolism of Mtb on glucose but not on fatty acids. Importantly, rv0158 KO exhibited enhanced growth on propionate, and the Rv0158 protein directly binds to methylmalonyl-CoA, a key intermediate in propionate catabolism. Metabolite profiling, ChIP-Seq, and gene-expression analyses indicate that Rv0158 manages metabolic neutralization of propionate toxicity by regulating the methylcitrate cycle. Disruption of rv0158 enhanced the sensitivity of Mtb to oxidative stress, nitric oxide, and anti-TB drugs. Lastly, rv0158 KO showed poor survival in macrophages and persistence defect in mice. Our results suggest that Rv0158 is a metabolic integrator for carbon metabolism and redox balance in Mtb.

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