1. 3 minutes to precisely measure morphogen concentration
- Author
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Cécile Fradin, Aleksandra M. Walczak, Nathalie Dostatni, Carmina Angelica Perez Romero, Aurelien Guillou, Huy Tran, Tanguy Lucas, Mathieu Coppey, Dynamique du noyau [Institut Curie], Institut Curie [Paris]-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire de Physique Théorique de l'ENS (LPTENS), Fédération de recherche du Département de physique de l'Ecole Normale Supérieure - ENS Paris (FRDPENS), Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS Paris), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), Laboratoire Physico-Chimie Curie [Institut Curie] (PCC), Institut Curie [Paris]-Institut de Chimie du CNRS (INC)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS), HAL UPMC, Gestionnaire, Laboratoire de Physique Théorique de l'ENS [École Normale Supérieure] (LPTENS), École normale supérieure - Paris (ENS-PSL), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-École normale supérieure - Paris (ENS-PSL), and Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Centre National de la Recherche Scientifique (CNRS)-Sorbonne Université (SU)-Centre National de la Recherche Scientifique (CNRS)
- Subjects
0301 basic medicine ,Embryology ,Cancer Research ,Embryo, Nonmammalian ,Zygote ,Gene Expression ,Cooperativity ,Biochemistry ,Database and Informatics Methods ,0302 clinical medicine ,Transcription (biology) ,Morphogenesis ,Drosophila Proteins ,Cell Cycle and Cell Division ,Promoter Regions, Genetic ,[SDV.BDD]Life Sciences [q-bio]/Development Biology ,Genetics (clinical) ,0303 health sciences ,biology ,Chromosome Biology ,Chemistry ,Transcriptional Control ,Optical Imaging ,Gene Expression Regulation, Developmental ,Drosophila embryogenesis ,Cell biology ,Drosophila melanogaster ,Cell Processes ,embryonic structures ,[SDV.BBM.GTP] Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,Sequence Analysis ,Research Article ,Morphogen ,animal structures ,lcsh:QH426-470 ,Bioinformatics ,DNA transcription ,Mitosis ,Research and Analysis Methods ,03 medical and health sciences ,Sequence Motif Analysis ,Live cell imaging ,[SDV.BBM.GTP]Life Sciences [q-bio]/Biochemistry, Molecular Biology/Genomics [q-bio.GN] ,[SDV.BDD] Life Sciences [q-bio]/Development Biology ,DNA-binding proteins ,Genetics ,Animals ,Gene Regulation ,Molecular Biology ,Transcription factor ,Ecology, Evolution, Behavior and Systematics ,Body Patterning ,030304 developmental biology ,Homeodomain Proteins ,Binding Sites ,Embryos ,Biology and Life Sciences ,Proteins ,Promoter ,Cell Biology ,Molecular Development ,biology.organism_classification ,Regulatory Proteins ,Morphogens ,lcsh:Genetics ,030104 developmental biology ,Trans-Activators ,Developmental biology ,030217 neurology & neurosurgery ,Developmental Biology ,Transcription Factors - Abstract
Morphogen gradients provide concentration-dependent positional information along polarity axes. Although the dynamics of the establishment of these gradients is well described, precision and noise in the downstream activation processes remain elusive. A simple paradigm to address these questions is the Bicoid morphogen gradient that elicits a rapid step-like transcriptional response in young fruit fly embryos. Focusing on the expression of the major Bicoid target, hunchback (hb), at the onset of zygotic transcription, we used the MS2-MCP approach which combines fluorescent labeling of nascent mRNA with live imaging at high spatial and temporal resolution. Removing 36 putative Zelda binding sites unexpectedly present in the original MS2 reporter, we show that the 750 bp of the hb promoter are sufficient to recapitulate endogenous expression at the onset of zygotic transcription. After each mitosis, in the anterior, expression is turned on to rapidly reach a plateau with all nuclei expressing the reporter. Consistent with a Bicoid dose-dependent activation process, the time period required to reach the plateau increases with the distance to the anterior pole. Despite the challenge imposed by frequent mitoses and high nuclei-to-nuclei variability in transcription kinetics, it only takes 3 minutes at each interphase for the MS2 reporter loci to distinguish subtle differences in Bicoid concentration and establish a steadily positioned and steep (Hill coefficient ~ 7) expression boundary. Modeling based on the cooperativity between the 6 known Bicoid binding sites in the hb promoter region, assuming rate limiting concentrations of the Bicoid transcription factor at the boundary, is able to capture the observed dynamics of pattern establishment but not the steepness of the boundary. This suggests that a simple model based only on the cooperative binding of Bicoid is not sufficient to describe the spatiotemporal dynamics of early hb expression., Author summary During development, the first thing that an embryo needs to know is the orientation of its body. We study how the head-to-tail axis forms in the fruit fly embryo. To position the axis, the embryo relies on proteins called morphogens broadcasting instructions to other genes, so that cells know, depending on where they are along the axis, what they should become. The concentration of the Bicoid morphogen is much higher in the head of the embryo and lower towards the tail. Bicoid activates hunchback, which divides the embryo in two parts. By visualizing this process in real time in living embryos, we see the hunchback gene activated very efficiently in the head part, where each nucleus expresses the gene, whereas it is not expressed at all in the tail part. Intriguingly, the boundary separating the two domains is precisely positioned and becomes very steep in no more than three minutes. We use a modeling approach to understand how this is achieved so rapidly. Given the parameters of the system, we find that although our model is able to reproduce the fast dynamics of the process, it fails to reproduce the steepness of the boundary suggesting that a more complex approach is needed to capture the additional mechanisms involved.
- Published
- 2018
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