Academic literature on the topic 'Bicoid-Hunchback'

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Journal articles on the topic "Bicoid-Hunchback"

1

Janody, F., J. Reischl, and N. Dostatni. "Persistence of Hunchback in the terminal region of the Drosophila blastoderm embryo impairs anterior development." Development 127, no. 8 (April 15, 2000): 1573–82. http://dx.doi.org/10.1242/dev.127.8.1573.

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Anterior terminal development is controlled by several zygotic genes that are positively regulated at the anterior pole of Drosophila blastoderm embryos by the anterior (bicoid) and the terminal (torso) maternal determinants. Most Bicoid target genes, however, are first expressed at syncitial blastoderm as anterior caps, which retract from the anterior pole upon activation of Torso. To better understand the interaction between Bicoid and Torso, a derivative of the Gal4/UAS system was used to selectively express the best characterised Bicoid target gene, hunchback, at the anterior pole when its expression should be repressed by Torso. Persistence of hunchback at the pole mimics most of the torso phenotype and leads to repression at early stages of a labral (cap'n'collar) and two foregut (wingless and hedgehog) determinants that are positively controlled by bicoid and torso. These results uncovered an antagonism between hunchback and bicoid at the anterior pole, whereas the two genes are known to act in concert for most anterior segmented development. They suggest that the repression of hunchback by torso is required to prevent this antagonism and to promote anterior terminal development, depending mostly on bicoid activity.
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Wolff, C., R. Schroder, C. Schulz, D. Tautz, and M. Klingler. "Regulation of the Tribolium homologues of caudal and hunchback in Drosophila: evidence for maternal gradient systems in a short germ embryo." Development 125, no. 18 (September 15, 1998): 3645–54. http://dx.doi.org/10.1242/dev.125.18.3645.

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In short germ embryos, the germ rudiment forms at the posterior ventral side of the egg, while the anterior-dorsal region becomes the extra-embryonic serosa. It is difficult to see how an anterior gradient like that of bicoid in Drosophila could in these embryos be directly involved in patterning of the germ rudiment. Moreover, since it has not yet been possible to recover a bicoid homologue from any species outside the diptera, it has been speculated that the anterior bicoid gradient could be a late addition during insect evolution. We addressed this question by analysing the regulation of potential target genes of bicoid in the short germ embryo of Tribolium castaneum. We demonstrate that homologues of caudal and hunchback from Tribolium are regulated by Drosophila bicoid. In Drosophila, maternal caudal RNA is translationally repressed by bicoid. We find that Tribolium caudal RNA is also translationally repressed by bicoid, when it is transferred into Drosophila embryos under a maternal promoter. This strongly suggests that a functional bicoid homologue must exist in Tribolium. The second target gene, hunchback, is transcriptionally activated by bicoid in Drosophila. Transfer of the regulatory region of Tribolium hunchback into Drosophila also results in regulation by early maternal factors, including bicoid, but in a pattern that is more reminiscent of Tribolium hunchback expression, namely in two early blastoderm domains. Using enhancer mapping constructs and footprinting, we show that caudal activates the posterior of these domains via a specific promoter. Our experiments suggest that a major event in the evolutionary transition from short to long germ embryogenesis was the switch from activation of the hunchback gap domain by caudal to direct activation by bicoid. This regulatory switch can explain how this domain shifted from a posterior location in short germ embryos to its anterior position in long germ insects, and it also suggest how an anterior gradient can pattern the germ rudiment in short germ embryos, i.e. by regulating the expression of caudal.
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Ma, X., D. Yuan, K. Diepold, T. Scarborough, and J. Ma. "The Drosophila morphogenetic protein Bicoid binds DNA cooperatively." Development 122, no. 4 (April 1, 1996): 1195–206. http://dx.doi.org/10.1242/dev.122.4.1195.

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The Drosophila morphogenetic protein Bicoid, encoded by the maternal gene bicoid, is required for the development of the anterior structures in the embryo. Bicoid, a transcriptional activator containing a homeodomain, is distributed in an anterior-to-posterior gradient in the embryo. In response to this gradient, the zygotic gene hunchback is expressed uniformly in the anterior half of the embryo in a nearly all-or-none manner. In this report we demonstrate that a recombinant Bicoid protein binds cooperatively to its sites within a hunchback enhancer element. A less than 4-fold increase in Bicoid concentration is sufficient to achieve an unbound/bound transition in DNA binding. Using various biochemical and genetic methods we further demonstrate that Bicoid molecules can interact with each other. Our results are consistent with previous studies performed in the embryo, and they suggest that one mechanism to achieve a sharp on/off switch of gene expression in response to a morphogenetic gradient is cooperative DNA binding facilitated by protein-protein interaction.
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Schulz, C., and D. Tautz. "Autonomous concentration-dependent activation and repression of Kruppel by hunchback in the Drosophila embryo." Development 120, no. 10 (October 1, 1994): 3043–49. http://dx.doi.org/10.1242/dev.120.10.3043.

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The subdivision of the anterior-posterior axis in Drosophila is achieved by a cascade of spatially regulated transcription factors which form short-range gradients at the syncytial blastoderm stage. These factors are assumed to have concentration-dependent regulatory effects on their target genes. However, there is so far little direct in vivo evidence that a single factor can autonomously activate and repress a given target gene. We have analysed here the regulatory capabilities of the gap gene hunchback by creating an artificial gradient of hunchback in the early embryo. This was achieved by providing the maternally expressed mRNA of hunchback with the anterior localization signal of the bicoid RNA. The effects of this artificial hunchback gradient were then studied in different types of mutant background. We show that under these conditions hb is autonomously capable of activating the target gene Kruppel at low concentrations and repressing it at high concentrations. In addition, we show that the artificially created hunchback gradient can organize a large part of the segment pattern, although it is expressed at a different position and in a different shape than the wild-type gradient of hunchback.
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Gaul, U., and H. Jackle. "Analysis of maternal effect mutant combinations elucidates regulation and function of the overlap of hunchback and Kruppel gene expression in the Drosophila blastoderm embryo." Development 107, no. 3 (November 1, 1989): 651–62. http://dx.doi.org/10.1242/dev.107.3.651.

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The metameric organisation of the Drosophila embryo is generated early during development, due to the action of maternal effect and zygotic segmentation and homeotic genes. The gap genes participate in the complex process of pattern formation by providing a link between the maternal and the zygotic gene activities. Under the influence of maternal gene products they become expressed in distinct domains along the anteroposterior axis of the embryo; negative interactions between neighboring gap genes are thought to be involved in establishing the expression domains. The gap gene activities in turn are required for the correct patterning of the pair-rule genes; little is known, however, about the underlying mechanisms. We have monitored the distribution of gap and pair-rule genes in wild-type embryos and in embryos in which the anteroposterior body pattern is greatly simplified due to combinations of maternal effect mutations (staufen exuperantia, vasa exuperantia, vasa exuperantia, bicoid oskar, bicoid oskar torsolike, vasa torso exuperantia). We show that the domains of protein distribution of the gap genes hunchback and Kruppel overlap in wild-type embryos. Based on the analysis of the maternal mutant combinations, we suggest an explanation of how this overlap is generated. Furthermore, our data show that different constellations of gap gene activities provide different input for the pair-rule genes, and thus strongly suggest that the overlap of hunchback and Kruppel in wild-type is functional in the formation of the patterns of pair-rule genes.
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6

Sommer, R., and D. Tautz. "Segmentation gene expression in the housefly Musca domestica." Development 113, no. 2 (October 1, 1991): 419–30. http://dx.doi.org/10.1242/dev.113.2.419.

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Drosophila and Musca both belong to the group of higher dipteran flies and show morphologically a very similar early development. However, these two species are evolutionary separated by at least 100 million years. This presents the opportunity for a comparative analysis of segmentation gene expression across a large evolutionary distance in a very similar embryonic background. We have analysed in detail the early expression of the maternal gene bicoid, the gap genes hunchback, Kruppel, knirps and tailless, the pair-rule gene hairy, the segment-polarity gene engrailed and the homoeotic gene Ultrabithorax. We show that the primary expression domains of these genes are conserved, while some secondary expression aspects have diverged. Most notable is the finding of hunchback expression in 11–13 stripes shortly before gastrulation, as well as a delayed expression of terminal domains of various genes. We conclude that the early developmental gene hierarchy, as it has been defined in Drosophila, is evolutionary conserved in Musca domestica.
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7

Schröder, Reinhard. "The genes orthodenticle and hunchback substitute for bicoid in the beetle Tribolium." Nature 422, no. 6932 (April 2003): 621–25. http://dx.doi.org/10.1038/nature01536.

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8

Moudgil, Anshika, Ranbir Chander Sobti, and Tejinder Kaur. "In-silico identification and comparison of transcription factor binding sites cluster in anterior-posterior patterning genes in Drosophila melanogaster and Tribolium castaneum." PLOS ONE 18, no. 8 (August 17, 2023): e0290035. http://dx.doi.org/10.1371/journal.pone.0290035.

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The cis-regulatory data that help in transcriptional regulation is arranged into modular pieces of a few hundred base pairs called CRMs (cis-regulatory modules) and numerous binding sites for multiple transcription factors are prominent characteristics of these cis-regulatory modules. The present study was designed to localize transcription factor binding site (TFBS) clusters on twelve Anterior-posterior (A-P) genes in Tribolium castaneum and compare them to their orthologous gene enhancers in Drosophila melanogaster. Out of the twelve A-P patterning genes, six were gap genes (Kruppel, Knirps, Tailless, Hunchback, Giant, and Caudal) and six were pair rule genes (Hairy, Runt, Even-skipped, Fushi-tarazu, Paired, and Odd-skipped). The genes along with 20 kb upstream and downstream regions were scanned for TFBS clusters using the Motif Cluster Alignment Search Tool (MCAST), a bioinformatics tool that looks for set of nucleotide sequences for statistically significant clusters of non-overlapping occurrence of a given set of motifs. The motifs used in the current study were Hunchback, Caudal, Giant, Kruppel, Knirps, and Even-skipped. The results of the MCAST analysis revealed the maximum number of TFBS for Hunchback, Knirps, Caudal, and Kruppel in both D. melanogaster and T. castaneum, while Bicoid TFBS clusters were found only in D. melanogaster. The size of all the predicted TFBS clusters was less than 1kb in both insect species. These sequences revealed more transversional sites (Tv) than transitional sites (Ti) and the average Ti/Tv ratio was 0.75.
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Stauber, M., H. Taubert, and U. Schmidt-Ott. "Function of bicoid and hunchback homologs in the basal cyclorrhaphan fly Megaselia (Phoridae)." Proceedings of the National Academy of Sciences 97, no. 20 (September 19, 2000): 10844–49. http://dx.doi.org/10.1073/pnas.190095397.

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Bonneton, François, Philip J. Shaw, Claire Fazakerley, Min Shi, and Gabriel A. Dover. "Comparison of bicoid-dependent regulation of hunchback between Musca domestica and Drosophila melanogaster." Mechanisms of Development 66, no. 1-2 (August 1997): 143–56. http://dx.doi.org/10.1016/s0925-4773(97)00100-7.

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Dissertations / Theses on the topic "Bicoid-Hunchback"

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Crauk, Olivier. "Exploration moléculaire de l' activité du gradient morphogénétique Bicoid dans l' embryon précoce de drosophile." Paris 6, 2005. http://www.theses.fr/2005PA066285.

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Schaeffer, Valérie. "Rôle de Bicoid chez la drosophile : intercations avec Hunchback et Torso." Montpellier 2, 1999. http://www.theses.fr/1999MON20102.

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Lors du developpement precoce de la drosophile, le plan de l'embryon est defini par quatre systemes maternels. La molecule-cle du systeme anterieur est bicoid, facteur de transcription a homeodomaine. Il a ete montre que bicoid etait aide par un autre morphogene anterieur, hunchback. Bod et hb sont capables d'agir en synergie pour organiser un developpement anterieur correct. Le mecanisme moleculaire propose pour cette synergie fait intervenir des facteurs specifiques de la machinerie generale de transcription, les tafs. Dans un premier temps, nous avons ete amenes a tester la signification biologique des ces resultats in vivo. Des mouches transgeniques portant des deletions de la proteine bicoid ont montre que les domaines putatifs d'activation de la transcription de bicoid etaient dispensables pour son activite in vivo. Ceci suggere que les interactions observees in vitro avec des composants specifiques de la machinerie generale de transcription ne sont pas essentiels pour un developpement anterieur correct. Le systeme terminal anterieur a longtemps ete implique dans la modification de l'activite de bcd. A l'extremite la plus anterieure, tor est connu pour avoir une fonction negative sur l'activite de bcd (retraction des genes cibles de bcd) tandis que dans les regions posterieures de la tete presomptive, tor semble plutot augmenter la fonction de bcd. Dans une deuxieme partie, nous montrons que de multiples copies de bcd sont capables de restaurer le phenotype anterieur des mutants du systeme terminal. Ceci signifie que le seul role de tor dans l'anterieur est d'aider bcd a effectuer sa fonction. Nous avons aussi montres que tor n'agit pas sur bcd mais qu'il existe deux voies d'activation differentes des genes cibles de tor et bcd. Ces observations sont consistantes avec l'hypothese de l'apparition recente d'un centre morphogenetique anterieur (bcd et anterieur tor) chez les dipteres superieurs.
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3

Lucas, Tanguy. "Spatio-temporal regulation of hunchback during the zygotic genome activation in Drosophila." Thesis, Paris 6, 2015. http://www.theses.fr/2015PA066707.

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Les gradients morphogénétiques contrôlent l'émergence de polarités axiales au cours du développement. Bien que la dynamique d'établissement de ces gradients soit bien comprise, la précision des mécanismes d'activation agissant en aval restent à élucider. Nous abordons cette question avec le gradient de Bicoid qui fournit rapidement une réponse transcriptionnelle robuste dans l'embryon de drosophile. Cette robustesse survient malgré le challenge imposé par de fréquentes mitoses dépourvues de transcription. Un calcul théorique intégrant les paramètres physiques de Bicoid (concentration, diffusion) indique que la mesure précise de concentration de Bicoid ne peut être effectuée à chaque interphase en 5-6mn. Il a donc été proposé que l'acquisition rapide de cette robustesse repose sur une mémorisation de l'état transcriptionnel au cours des divisions. Pour tester cette hypothèse, j'ai adapté à l'embryon de drosophile le système MS2 d'étiquetage des ARN dans les cellules vivantes et démontré qu'il permettait de suivre la dynamique transcriptionnelle dans un organisme pluricellulaire vivant. De manière inattendue, le rapporteur MS2 s'exprime aussi postérieurement ce qui m'a empêché de tester l'hypothèse de mémorisation. J'ai montré que cette expression postérieure est due à la présence de sites de fixation pour le facteur de transcription Zelda dans la cassette MS2. Un nouveau rapporteur MS2, dépourvu de ces sites récapitule l'expression endogène et fournit un outil de choix pour tester l'hypothèse de mémorisation. Ce travail ouvre de nouvelles perspectives pour comprendre la dynamique transcriptionnelle sur laquelle repose l'émergence des patrons d'expression développementaux
Morphogen gradients provide concentration-dependent positional information along polarity axes. Although the dynamics of these gradients is well described, precision and noise in the activation processes acting downstream remain unclear. To address this question, we study the response to the Bicoid gradient that elicits very rapidly a robust transcriptional response in young fly embryos. This robustness occurs despite the challenge imposed by frequent mitoses during which transcription is interrupted suggesting that nuclei measure the Bicoid concentration during the 5-6 mn interphases. Modeling using statistical mechanics and Bicoid physical parameters do not account for accurate measurement of Bicoid concentration in such a short period. It was proposed that rapid robustness of the Bicoid response relies on a memorization process allowing nuclei to recall Bicoid concentration from previous cycles. To understand how the Bicoid system resists to the challenge imposed by mitosis, I have adapted the MS2 RNA-tagging approach to fly embryos and shown that it can be used to quantify transcription dynamics in a living multicellular organism. Unexpectedly, the MS2 reporter was also expressed in the posterior of the embryo making it impossible to directly test the memorization hypothesis. I have shown that this posterior expression is due to binding sites for the transcription factor Zelda unexpectedly localized in the MS2 cassette. A newly engineered MS2 reporter removing those sites faithfully reproduces the endogenous expression providing a powerful tool to test the memory hypothesis. This work opens new avenue to decipher the transcription dynamics underlying pattern formation
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Sasai, Masaki, Suguru Oho, Hiroki Murakami, and Yurie Okabe-Oho. "Stable, Precise, and Reproducible Patterning of Bicoid and Hunchback Molecules in the Early Drosophila Embryo." PLOS, 2009. http://hdl.handle.net/2237/20618.

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Shaw, Philip James. "Molecular characterisation of the interaction between the bicoid and hunchback genes in Musca domestica : insights into the evolution of a regulatory interaction." Thesis, University of Leicester, 1998. http://hdl.handle.net/2381/30312.

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Regulatory evolution of genes controlling development is widely considered to be the major driving force of morphological change in the evolution of animal lineages. The regulation of genes is controlled by interactions between trans-acting factors and cis-regulatory DNA. The evolution of the interaction between the bicoid and hunchback genes is studied by comparing this orthologous interaction in Drosophila melanogaster with the housefly, Musca domestica. A genomic clone of the hunchback region from Musca domestica from an earlier work was characterised by restriction mapping and sequencing. A promoter region upstream of an early embryonically expressed transcript was mapped to the hunchback genomic sequence. Sites of specific interaction between a bacterially-derived Musca domestica BICOID homeodomain protein were mapped to the promoter by DNaseI footprinting. The hunchback regulatory sequence of Musca domestica are unalignable with Drosophila melanogaster; nevertheless, similar patterns of repetitive sequence distribution were found by computer-aided analysis, which suggest extensive slippage-like processes may have been important in the evolution of the hunchback region. A promoter element of Musca domestica hunchback drives a bicoid-dependent pattern of expression in the anterior of transgenic Drosophila melanogaster embryos. A later ectopic expression points to a functional divergence of promoter recognition between the two species. The in vitro DNA-binding function of the Musca domestica BICOID homeodomain was further characterised by quantitative gel-shift assays. The data suggest that the homeodomain differences between Musca domestica and Drosophila melanogaster reflect differences in sequence recognition. The homeodomain differences may have co-evolved with the binding site sequences in the hunchback promoters. It is argued that molecular coevolution of the BICOID trans-factor and cis-binding sites in the hunchback promoter has accompanied a divergence in embryonic size between Musca domestica and Drosophila melanogaster.
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6

Lucas, Tanguy. "Spatio-temporal regulation of hunchback during the zygotic genome activation in Drosophila." Electronic Thesis or Diss., Paris 6, 2015. http://www.theses.fr/2015PA066707.

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Les gradients morphogénétiques contrôlent l'émergence de polarités axiales au cours du développement. Bien que la dynamique d'établissement de ces gradients soit bien comprise, la précision des mécanismes d'activation agissant en aval restent à élucider. Nous abordons cette question avec le gradient de Bicoid qui fournit rapidement une réponse transcriptionnelle robuste dans l'embryon de drosophile. Cette robustesse survient malgré le challenge imposé par de fréquentes mitoses dépourvues de transcription. Un calcul théorique intégrant les paramètres physiques de Bicoid (concentration, diffusion) indique que la mesure précise de concentration de Bicoid ne peut être effectuée à chaque interphase en 5-6mn. Il a donc été proposé que l'acquisition rapide de cette robustesse repose sur une mémorisation de l'état transcriptionnel au cours des divisions. Pour tester cette hypothèse, j'ai adapté à l'embryon de drosophile le système MS2 d'étiquetage des ARN dans les cellules vivantes et démontré qu'il permettait de suivre la dynamique transcriptionnelle dans un organisme pluricellulaire vivant. De manière inattendue, le rapporteur MS2 s'exprime aussi postérieurement ce qui m'a empêché de tester l'hypothèse de mémorisation. J'ai montré que cette expression postérieure est due à la présence de sites de fixation pour le facteur de transcription Zelda dans la cassette MS2. Un nouveau rapporteur MS2, dépourvu de ces sites récapitule l'expression endogène et fournit un outil de choix pour tester l'hypothèse de mémorisation. Ce travail ouvre de nouvelles perspectives pour comprendre la dynamique transcriptionnelle sur laquelle repose l'émergence des patrons d'expression développementaux
Morphogen gradients provide concentration-dependent positional information along polarity axes. Although the dynamics of these gradients is well described, precision and noise in the activation processes acting downstream remain unclear. To address this question, we study the response to the Bicoid gradient that elicits very rapidly a robust transcriptional response in young fly embryos. This robustness occurs despite the challenge imposed by frequent mitoses during which transcription is interrupted suggesting that nuclei measure the Bicoid concentration during the 5-6 mn interphases. Modeling using statistical mechanics and Bicoid physical parameters do not account for accurate measurement of Bicoid concentration in such a short period. It was proposed that rapid robustness of the Bicoid response relies on a memorization process allowing nuclei to recall Bicoid concentration from previous cycles. To understand how the Bicoid system resists to the challenge imposed by mitosis, I have adapted the MS2 RNA-tagging approach to fly embryos and shown that it can be used to quantify transcription dynamics in a living multicellular organism. Unexpectedly, the MS2 reporter was also expressed in the posterior of the embryo making it impossible to directly test the memorization hypothesis. I have shown that this posterior expression is due to binding sites for the transcription factor Zelda unexpectedly localized in the MS2 cassette. A newly engineered MS2 reporter removing those sites faithfully reproduces the endogenous expression providing a powerful tool to test the memory hypothesis. This work opens new avenue to decipher the transcription dynamics underlying pattern formation
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7

Fernandes, Gonçalo. "Imaging transcription in living embryos to decipher the robustness of patterning." Electronic Thesis or Diss., Université Paris sciences et lettres, 2022. http://www.theses.fr/2022UPSLS025.

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Abstract:
Au cours du développement, les gradients morphogénétiques fournissent l’information positionnelle nécessaire à l’établissement des axes de polarité. Si l’importance de ces gradients est bien établie, la façon dont ils assurent la reproductibilité des patrons d’expression malgré la nature stochastique de la transcription reste énigmatique.Pour répondre à cette question, j’étudie l’établissement de l’axe antéro-postérieur (AP) de l’embryon de drosophile, sous contrôle de Bicoid (Bcd), un facteur de transcription et morphogène bien connu. Les ARNs de bcd sont exprimés maternellement et ancrés au pôle antérieur de l’ovocyte. Après la ponte, ces ARNs sont traduits en protéines, qui diffusent dans le cytoplasme pour former un gradient de concentration exponentiel avec son maximum au pôle antérieur. Un modèle simple propose que le destin de chaque cellule le long de l’axe est déterminé par la concentration de Bcd qu’elle reçoit. Toutefois, de nombreux débats ont remis en question la possibilité que les patrons d’expression induits par Bcd résultent uniquement des propriétés de diffusion et d’interaction des protéines Bcd avec leurs séquences cibles.L’objectif de ma thèse était de comprendre le rôle précis de Bcd dans l’expression de son gène cible principal, hunchback (hb). Pour cela, j’ai adapté à des gènes rapporteurs synthétiques, le système MS2-MCP, qui permet l’étiquetage fluorescent des ARN et l’analyse quantitative de la dynamique transcriptionnelle avec une forte résolution spatio-temporelle dans les embryons vivants. Dans ces rapporteurs, la séquence MS2 est contrôlée par un promoteur minimal contenant aussi les sites de fixation pour Bcd ou ses partenaires, Hb et Zelda (Zld), seul ou en combinaison. Mon but était de déterminer comment les divers rapporteurs récapitulent l’expression du promoteur d’hb et d’identifier les rôles spécifiques de chacun de ces facteurs et leurs interactions dans le mécanisme de transcription.De façon intéressante, l’expression du rapporteur avec uniquement neuf sites pour Bcd (trois de plus que dans le gène hb) récapitule l’expression du rapporteur hb-MS2, mais de façon moins rapide et avec un domaine d’expression à la bordure moins nette. Cela suggère qu’en définissant la position de la bordure mais pas sa netteté ni la rapidité de son établissement, Bcd est la source principale d’information positionnelle.En outre, la fixation des partenaires de Bcd au promoteur accélère le processus en agissant à différentes étapes : i) Hb agit en synergie avec Bcd en réduisant les fluctuations du promoteur et en augmentant le taux de démarrage de la polymérase ; ii) Zld abaisse le seuil de concentration de Bcd nécessaire à l’activation par Bcd. En collaboration avec des physiciens, nous avons développé un modèle de transcription impliquant Bcd et fournissant un contexte théorique aux données expérimentales. Ce modèle montre que l’établissement rapide de la bordure d’expression d’hb peut être expliqué par une réaction d’équilibre entre Bcd et ses sites de fixation pour l’information positionnelle nécessitant Zld et Hb pour sa dynamique temporelle.Pour confirmer que Bcd est la seule source d’information positionnelle du système, j’ai comparé la position de la bordure d’expression des gènes rapporteurs dépendant uniquement de Bcd dans des embryons exprimant une ou une demi-dose de Bcd. De manière surprenante, pour chaque rapporteur, le déplacement de la bordure est plus faible que son estimation théorique tenant compte de la constante du gradient exponentiel de concentration de Bcd. Cela indique une constante plus faible pour le gradient d’activité de Bcd et suggère l’existence de sous-populations de Bcd, avec certaines molécules moins actives que d’autres. L’amplitude du déplacement de la bordure du gène rapporteur hb-MS2 est la même que celle obtenue pour les rapporteurs dépendant uniquement de Bcd ce qui confirme que Bcd est bien la seule source d’information positionnelle pour l’expression d’hb
Morphogen gradients provide concentration-dependent positional information required to establish the polarity of developmental axes. Although the critical role of these gradients is well recognized, it is unclear how they provide reproducible expression patterns. This is particularly surprising if we consider the stochastic nature of transcription.To address this question, I focus on the establishment of the anterior-posterior (AP) axis of fruit fly embryos, which is mostly defined by Bicoid (Bcd), a very well-characterized morphogen and transcription factor. bcd mRNAs are expressed maternally and anchored at the anterior tip of the oocyte. After egg laying, these mRNAs are translated into proteins, which diffuse through the cytoplasm and form a gradient with its highest concentration at the anterior. A simple model is that depending on their position along the AP axis and thus on Bcd concentration, cells will adopt different fates. However, long debates in the field have questioned the possibility that Bcd-dependent transcription patterns emerge solely from diffusive biochemical interactions between limiting amounts of Bcd molecules and the gene promoter region.The goal of my PhD was to determine how Bcd precisely regulates expression of its main target gene, hunchback (hb). For this, I adapted to synthetic reporters the MS2-MCP system, which allows the fluorescent tagging of mRNAs and provides, thus, a quantitative analysis of transcription dynamics at high spatiotemporal resolution in living embryos. In these reporters, the MS2 sequence was placed under the control of a minimal promoter also containing DNA binding sites for Bcd and/or its known partners, Hb and Zelda (Zld), either alone or in combination. My goal was to determine how the various reporters could recapitulate expression of the hb promoter (hb-MS2 reporter) and shed light on the specific roles of the different factors and their interactions in the transcription mechanism.Interestingly, expression of the reporter with only nine Bcd binding sites (three more than in the hb gene) matches almost perfectly the hb-MS2 reporter pattern, except for the very high steepness of the expression domain boundary and the speed to reach steady-state. This suggests that Bcd alone is the main source of positional information, defining the positioning of the boundary but not its steepness nor the speed of its establishment.In addition, binding of Bcd’s partners to the promoter speed-up the process by acting in different steps of the transcription mechanism: i) Hb synergizes with Bcd by reducing transcription burstiness and increasing the polymerase firing rate; ii) Zld lowers the Bcd concentration threshold required for Bcd-dependent expression. In collaboration with physicists, a biophysical model of Bcd-dependent expression was developed providing a theoretical framework for the experimental data. This model showed that the very rapid establishment of the hb expression boundary can be solely explained by an equilibrium model involving the binding of Bcd molecules to their DNA-binding sites for positional information which requires Zld and Hb for its temporal dynamics.To further confirm that Bcd is the sole source of positional information for hb expression, I compared the boundary position of the Bcd-only dependent reporters in embryos expressing one dose or half dose of Bcd. Surprisingly, the corresponding shifts of these reporters’ boundaries upon one vs half dose of Bcd were smaller than theoretically expected given the measured decay length of the Bcd protein gradient. This indicates a shorter decay length for the Bcd activity gradient and suggests the existence of different Bcd populations, with some Bcd molecules being less active than others. Importantly, the shift observed for the hb-MS2 reporter was the same as for the Bcd-only dependent reporters confirming Bcd as the sole source of positional information for hb expression
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8

Lemke, Steffen Joachim. "Evolution of Bicoid-dependent hunchback Regulation in Diptera." Doctoral thesis, 2006. http://hdl.handle.net/11858/00-1735-0000-0006-B366-A.

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9

Lemke, Steffen [Verfasser]. "Evolution of bicoid-dependent hunchback regulation in diptera / submitted by Steffen Joachim Lemke." 2006. http://d-nb.info/987828843/34.

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