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1.
ArXiv ; 2024 Mar 05.
Article in English | MEDLINE | ID: mdl-38495568

ABSTRACT

Deciphering how genes interpret information from the concentration of transcription factors (TFs) within the cell nucleus remains a fundamental question in gene regulation. Recent advancements have unveiled the heterogeneous distribution of TF molecules in the nucleus, posing challenges to the precise decoding of concentration signals. To explore this phenomenon, we employ high-resolution single-cell imaging of a fluorescently tagged TF protein, Bicoid, in living fly embryos. We show that accumulation of Bicoid in submicron clusters preserves the spatial information of the maternal Bicoid gradient, and that cluster intensity, size, and frequency offer remarkably precise spatial cues. We further discover that various known gene targets of Bicoid activation colocalize with clusters and that for the target gene Hunchback, this colocalization is dependent on its enhancer binding affinity. Modeling information transfer through these clusters suggests that clustering offers a more rapid sensing mechanism for global nuclear concentrations than freely diffusing TF molecules detected by simple enhancers.

2.
Dev Cell ; 58(23): 2789-2801.e5, 2023 Dec 04.
Article in English | MEDLINE | ID: mdl-37890488

ABSTRACT

Transcription factor combinations play a key role in shaping cellular identity. However, the precise relationship between specific combinations and downstream effects remains elusive. Here, we investigate this relationship within the context of the Drosophila eve locus, which is controlled by gap genes. We measure spatiotemporal levels of four gap genes in heterozygous and homozygous gap mutant embryos and correlate them with the striped eve activity pattern. Although changes in gap gene expression extend beyond the manipulated gene, the spatial patterns of Eve expression closely mirror canonical activation levels in wild type. Interestingly, some combinations deviate from the wild-type repertoire but still drive eve activation. Although in homozygous mutants some Eve stripes exhibit partial penetrance, stripes consistently emerge at reproducible positions, even with varying gap gene levels. Our findings suggest a robust molecular canalization of cell fates in gap mutants and provide insights into the regulatory constraints governing multi-enhancer gene loci.


Subject(s)
Drosophila Proteins , Drosophila , Animals , Drosophila/metabolism , Gene Expression Regulation, Developmental/genetics , Transcription Factors/genetics , Transcription Factors/metabolism , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Gene Expression , Homeodomain Proteins/metabolism
3.
bioRxiv ; 2023 Dec 11.
Article in English | MEDLINE | ID: mdl-37398061

ABSTRACT

Shape changes of epithelia during animal development, such as convergent extension, are achieved through concerted mechanical activity of individual cells. While much is known about the corresponding large scale tissue flow and its genetic drivers, key open questions regard the cell-scale mechanics, e.g. internal vs external driving forces, and coordination, e.g. bottom-up self-organization vs top-down genetic instruction. To address these questions, we develop a quantitative, model-based analysis framework to relate cell geometry to local tension in recently obtained timelapse imaging data of gastrulating Drosophila embryos. This analysis provides a systematic decomposition of cell shape changes and T1-rearrangements into internally driven, active, and externally driven, passive, contributions. Specifically, we find evidence that germ band extension is driven by active T1 processes that self-organize through positive feedback acting on tensions. More generally, our findings suggest that epithelial convergent extension results from controlled transformation of internal force balance geometry which we quantify with a novel quantification tool for local tension configurations.

4.
Proc Natl Acad Sci U S A ; 119(15): e2112892119, 2022 04 12.
Article in English | MEDLINE | ID: mdl-35412853

ABSTRACT

During early Drosophila embryogenesis, a network of gene regulatory interactions orchestrates terminal patterning, playing a critical role in the subsequent formation of the gut. We utilized CRISPR gene editing at endogenous loci to create live reporters of transcription and light-sheet microscopy to monitor the individual components of the posterior gut patterning network across 90 min prior to gastrulation. We developed a computational approach for fusing imaging datasets of the individual components into a common multivariable trajectory. Data fusion revealed low intrinsic dimensionality of posterior patterning and cell fate specification in wild-type embryos. The simple structure that we uncovered allowed us to construct a model of interactions within the posterior patterning regulatory network and make testable predictions about its dynamics at the protein level. The presented data fusion strategy is a step toward establishing a unified framework that would explore how stochastic spatiotemporal signals give rise to highly reproducible morphogenetic outcomes.


Subject(s)
Body Patterning , Drosophila Proteins , Drosophila melanogaster , Endoderm , Gene Regulatory Networks , Animals , Body Patterning/genetics , Drosophila Proteins/genetics , Drosophila melanogaster/genetics , Drosophila melanogaster/growth & development , Endoderm/growth & development , Gene Expression Regulation, Developmental
5.
Cell Rep ; 38(12): 110543, 2022 03 22.
Article in English | MEDLINE | ID: mdl-35320726

ABSTRACT

Developmental patterning networks are regulated by multiple inputs and feedback connections that rapidly reshape gene expression, limiting the information that can be gained solely from slow genetic perturbations. Here we show that fast optogenetic stimuli, real-time transcriptional reporters, and a simplified genetic background can be combined to reveal the kinetics of gene expression downstream of a developmental transcription factor in vivo. We engineer light-controlled versions of the Bicoid transcription factor and study their effects on downstream gap genes in embryos. Our results recapitulate known relationships, including rapid Bicoid-dependent transcription of giant and hunchback and delayed repression of Krüppel. In addition, we find that the posterior pattern of knirps exhibits a quick but inverted response to Bicoid perturbation, suggesting a noncanonical role for Bicoid in directly suppressing knirps transcription. Acute modulation of transcription factor concentration while recording output gene activity represents a powerful approach for studying developmental gene networks in vivo.


Subject(s)
Drosophila Proteins , Homeodomain Proteins , Drosophila Proteins/metabolism , Gene Expression Regulation, Developmental , Homeodomain Proteins/metabolism , Optogenetics , Trans-Activators/metabolism
6.
Curr Biol ; 32(8): 1861-1868.e7, 2022 04 25.
Article in English | MEDLINE | ID: mdl-35290798

ABSTRACT

Gastrulation movements in all animal embryos start with regulated deformations of patterned epithelial sheets, which are driven by cell divisions, cell shape changes, and cell intercalations. Each of these behaviors has been associated with distinct aspects of gastrulation1-4 and has been a subject of intense research using genetic, cell biological, and more recently, biophysical approaches.5-14 Most of these studies, however, focus either on cellular processes driving gastrulation or on large-scale tissue deformations.15-23 Recent advances in microscopy and image processing create a unique opportunity for integrating these complementary viewpoints.24-28 Here, we take a step toward bridging these complementary strategies and deconstruct the early stages of gastrulation in the entire Drosophila embryo. Our approach relies on an integrated computational framework for cell segmentation and tracking and on efficient algorithms for event detection. The detected events are then mapped back onto the blastoderm shell, providing an intuitive visual means to examine complex cellular activity patterns within the context of their initial anatomic domains. By analyzing these maps, we identified that the loss of nearly half of surface cells to invaginations is compensated primarily by transient mitotic rounding. In addition, by analyzing mapped cell intercalation events, we derived direct quantitative relations between intercalation frequency and the rate of axis elongation. This work is setting the stage for systems-level dissection of a pivotal step in animal development.


Subject(s)
Embryo, Mammalian , Gastrulation , Animals , Cell Shape , Drosophila , Morphogenesis
7.
Proc Natl Acad Sci U S A ; 118(46)2021 11 16.
Article in English | MEDLINE | ID: mdl-34772813

ABSTRACT

In the regulation of gene expression, information of relevance to the organism is represented by the concentrations of transcription factor molecules. To extract this information the cell must effectively "measure" these concentrations, but there are physical limits to the precision of these measurements. We use the gap gene network in the early fly embryo as an example of the tradeoff between the precision of concentration measurements and the transmission of relevant information. For thresholded measurements we find that lower thresholds are more important, and fine tuning is not required for near-optimal information transmission. We then consider general sensors, constrained only by a limit on their information capacity, and find that thresholded sensors can approach true information theoretic optima. The information theoretic approach allows us to identify the optimal sensor for the entire gap gene network and to argue that the physical limitations of sensing necessitate the observed multiplicity of enhancer elements, with sensitivities to combinations rather than single transcription factors.


Subject(s)
Gene Regulatory Networks/genetics , Animals , Diptera/genetics , Gene Expression Regulation/genetics , Models, Biological , Transcription Factors/genetics
8.
Mol Syst Biol ; 17(8): e9895, 2021 08.
Article in English | MEDLINE | ID: mdl-34414660

ABSTRACT

The famous Arrhenius equation is well suited to describing the temperature dependence of chemical reactions but has also been used for complicated biological processes. Here, we evaluate how well the simple Arrhenius equation predicts complex multi-step biological processes, using frog and fruit fly embryogenesis as two canonical models. We find that the Arrhenius equation provides a good approximation for the temperature dependence of embryogenesis, even though individual developmental intervals scale differently with temperature. At low and high temperatures, however, we observed significant departures from idealized Arrhenius Law behavior. When we model multi-step reactions of idealized chemical networks, we are unable to generate comparable deviations from linearity. In contrast, we find the two enzymes GAPDH and ß-galactosidase show non-linearity in the Arrhenius plot similar to our observations of embryonic development. Thus, we find that complex embryonic development can be well approximated by the simple Arrhenius equation regardless of non-uniform developmental scaling and propose that the observed departure from this law likely results more from non-idealized individual steps rather than from the complexity of the system.


Subject(s)
Temperature
9.
Dev Biol ; 470: 147-153, 2021 02.
Article in English | MEDLINE | ID: mdl-33278404

ABSTRACT

The early stages of development involve complex sequences of morphological changes that are both reproducible from embryo to embryo and often robust to environmental variability. To investigate the relationship between reproducibility and robustness we examined cell cycle progression in early Drosophila embryos at different temperatures. Our experiments show that while the subdivision of cell cycle steps is conserved across a wide range of temperatures (5-35 â€‹°C), the relative duration of individual steps varies with temperature. We find that the transition into prometaphase is delayed at lower temperatures relative to other cell cycle events, arguing that it has a different mechanism of regulation. Using an in vivo biosensor, we quantified the ratio of activities of the major mitotic kinase, Cdk1 and one of the major mitotic phosphatases PP1. Comparing activation profile with cell cycle transition times at different temperatures indicates that in early fly embryos activation of Cdk1 drives entry into prometaphase but is not required for earlier cell cycle events. In fact, chromosome condensation can still occur when Cdk1 activity is inhibited pharmacologically. These results demonstrate that different kinases are rate-limiting for different steps of mitosis, arguing that robust inter-regulation may be needed for rapid and ordered mitosis.


Subject(s)
CDC2 Protein Kinase/metabolism , Cell Cycle Checkpoints , Cell Cycle , Drosophila Proteins/metabolism , Embryo, Nonmammalian/cytology , Mitosis , Animals , CDC2 Protein Kinase/antagonists & inhibitors , Cyclin B/metabolism , Drosophila Proteins/antagonists & inhibitors , Drosophila melanogaster/embryology , Embryo, Nonmammalian/enzymology , Enzyme Activation , Metaphase , Prometaphase , Prophase , Protein Phosphatase 1/metabolism , Temperature
10.
PLoS Comput Biol ; 16(8): e1008049, 2020 08.
Article in English | MEDLINE | ID: mdl-32822341

ABSTRACT

Tissue morphogenesis relies on repeated use of dynamic behaviors at the levels of intracellular structures, individual cells, and cell groups. Rapidly accumulating live imaging datasets make it increasingly important to formalize and automate the task of mapping recurrent dynamic behaviors (motifs), as it is done in speech recognition and other data mining applications. Here, we present a "template-based search" approach for accurate mapping of sub- to multi-cellular morphogenetic motifs using a time series data mining framework. We formulated the task of motif mapping as a subsequence matching problem and solved it using dynamic time warping, while relying on high throughput graph-theoretic algorithms for efficient exploration of the search space. This formulation allows our algorithm to accurately identify the complete duration of each instance and automatically label different stages throughout its progress, such as cell cycle phases during cell division. To illustrate our approach, we mapped cell intercalations during germband extension in the early Drosophila embryo. Our framework enabled statistical analysis of intercalary cell behaviors in wild-type and mutant embryos, comparison of temporal dynamics in contracting and growing junctions in different genotypes, and the identification of a novel mode of iterative cell intercalation. Our formulation of tissue morphogenesis using time series opens new avenues for systematic decomposition of tissue morphogenesis.


Subject(s)
Computational Biology/methods , Image Processing, Computer-Assisted/methods , Morphogenesis/physiology , Algorithms , Animals , Cell Division/physiology , Data Mining/methods , Drosophila/cytology , Drosophila/embryology , Embryo, Nonmammalian/cytology , Embryo, Nonmammalian/embryology , Female , Male , Microscopy, Confocal , Time Factors
11.
Dev Cell ; 52(6): 794-801.e4, 2020 03 23.
Article in English | MEDLINE | ID: mdl-32142631

ABSTRACT

Optogenetic perturbations, live imaging, and time-resolved ChIP-seq assays in Drosophila embryos were used to dissect the ERK-dependent control of the HMG-box repressor Capicua (Cic), which plays critical roles in development and is deregulated in human spinocerebellar ataxia and cancers. We established that Cic target genes are activated before significant downregulation of nuclear localization of Cic and demonstrated that their activation is preceded by fast dissociation of Cic from the regulatory DNA. We discovered that both Cic-DNA binding and repression are rapidly reinstated in the absence of ERK activation, revealing that inductive signaling must be sufficiently sustained to ensure robust transcriptional response. Our work provides a quantitative framework for the mechanistic analysis of dynamics and control of transcriptional repression in development.


Subject(s)
Drosophila Proteins/metabolism , HMGB Proteins/metabolism , MAP Kinase Signaling System , Repressor Proteins/metabolism , Active Transport, Cell Nucleus , Animals , Cell Nucleus/metabolism , Drosophila Proteins/genetics , Drosophila melanogaster , Extracellular Signal-Regulated MAP Kinases/metabolism , Gene Expression Regulation, Developmental , HMGB Proteins/genetics , Protein Binding , Repressor Proteins/genetics
12.
Cell ; 176(4): 844-855.e15, 2019 02 07.
Article in English | MEDLINE | ID: mdl-30712870

ABSTRACT

In developing organisms, spatially prescribed cell identities are thought to be determined by the expression levels of multiple genes. Quantitative tests of this idea, however, require a theoretical framework capable of exposing the rules and precision of cell specification over developmental time. We use the gap gene network in the early fly embryo as an example to show how expression levels of the four gap genes can be jointly decoded into an optimal specification of position with 1% accuracy. The decoder correctly predicts, with no free parameters, the dynamics of pair-rule expression patterns at different developmental time points and in various mutant backgrounds. Precise cellular identities are thus available at the earliest stages of development, contrasting the prevailing view of positional information being slowly refined across successive layers of the patterning network. Our results suggest that developmental enhancers closely approximate a mathematically optimal decoding strategy.


Subject(s)
GTPase-Activating Proteins/genetics , Gene Expression Regulation, Developmental/genetics , Gene Regulatory Networks/genetics , Animals , Body Patterning/genetics , Cell Differentiation/genetics , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Drosophila melanogaster/genetics , Drosophila melanogaster/metabolism , Embryo, Nonmammalian/metabolism , Embryonic Development/genetics , GTPase-Activating Proteins/metabolism , Gene Expression Regulation, Developmental/physiology , Models, Genetic , Transcription Factors/metabolism
13.
Elife ; 72018 02 09.
Article in English | MEDLINE | ID: mdl-29424685

ABSTRACT

During embryogenesis tissue layers undergo morphogenetic flow rearranging and folding into specific shapes. While developmental biology has identified key genes and local cellular processes, global coordination of tissue remodeling at the organ scale remains unclear. Here, we combine in toto light-sheet microscopy of the Drosophila embryo with quantitative analysis and physical modeling to relate cellular flow with the patterns of force generation during the gastrulation process. We find that the complex spatio-temporal flow pattern can be predicted from the measured meso-scale myosin density and anisotropy using a simple, effective viscous model of the tissue, achieving close to 90% accuracy with one time dependent and two constant parameters. Our analysis uncovers the importance of a) spatial modulation of myosin distribution on the scale of the embryo and b) the non-locality of its effect due to mechanical interaction of cells, demonstrating the need for the global perspective in the study of morphogenetic flow.


Subject(s)
Drosophila/embryology , Gastrulation , Myosins/analysis , Animals , Microscopy , Spatio-Temporal Analysis
14.
Elife ; 62017 09 11.
Article in English | MEDLINE | ID: mdl-28891464

ABSTRACT

In Drosophila, graded expression of the maternal transcription factor Bicoid (Bcd) provides positional information to activate target genes at different positions along the anterior-posterior axis. We have measured the genome-wide binding profile of Bcd using ChIP-seq in embryos expressing single, uniform levels of Bcd protein, and grouped Bcd-bound targets into four classes based on occupancy at different concentrations. By measuring the biochemical affinity of target enhancers in these classes in vitro and genome-wide chromatin accessibility by ATAC-seq, we found that the occupancy of target sequences by Bcd is not primarily determined by Bcd binding sites, but by chromatin context. Bcd drives an open chromatin state at a subset of its targets. Our data support a model where Bcd influences chromatin structure to gain access to concentration-sensitive targets at high concentrations, while concentration-insensitive targets are found in more accessible chromatin and are bound at low concentrations. This may be a common property of developmental transcription factors that must gain early access to their target enhancers while the chromatin state of the genome is being remodeled during large-scale transitions in the gene regulatory landscape.


Subject(s)
Chromatin/metabolism , Drosophila/embryology , Gene Expression Regulation, Developmental , Homeodomain Proteins/metabolism , Trans-Activators/metabolism , Animals , Chromatin Immunoprecipitation , Drosophila Proteins , Protein Binding , Sequence Analysis, DNA
15.
Proc Natl Acad Sci U S A ; 114(6): 1335-1340, 2017 02 07.
Article in English | MEDLINE | ID: mdl-28115706

ABSTRACT

Membraneless organelles play a central role in the organization of protoplasm by concentrating macromolecules, which allows efficient cellular processes. Recent studies have shown that, in vitro, certain components in such organelles can assemble through phase separation. Inside the cell, however, such organelles are multicomponent, with numerous intermolecular interactions that can potentially affect the demixing properties of individual components. In addition, the organelles themselves are inherently active, and it is not clear how the active, energy-consuming processes that occur constantly within such organelles affect the phase separation behavior of the constituent macromolecules. Here, we examine the phase separation model for the formation of membraneless organelles in vivo by assessing the two features that collectively distinguish it from active assembly, namely temperature dependence and reversibility. We use a microfluidic device that allows accurate and rapid manipulation of temperature and examine the quantitative dynamics by which six different nucleolar proteins assemble into the nucleoli of Drosophila melanogaster embryos. Our results indicate that, although phase separation is the main mode of recruitment for four of the studied proteins, the assembly of the other two is irreversible and enhanced at higher temperatures, behaviors indicative of active recruitment to the nucleolus. These two subsets of components differ in their requirements for ribosomal DNA; the two actively assembling components fail to assemble in the absence of ribosomal DNA, whereas the thermodynamically driven components assemble but lose temporal and spatial precision.


Subject(s)
Cell Nucleolus/metabolism , Drosophila Proteins/metabolism , Nuclear Proteins/chemistry , Animals , Animals, Genetically Modified , Drosophila melanogaster/genetics , Drosophila melanogaster/metabolism , Lab-On-A-Chip Devices , Thermodynamics
16.
Dev Biol ; 422(2): 125-134, 2017 02 15.
Article in English | MEDLINE | ID: mdl-28063874

ABSTRACT

The polarity protein Par3/Bazooka (Baz) has been established as a central component of the apical basal polarity system that determines the position of cell-cell junctions in epithelial cells. Consistent with that view, we show that shortly before gastrulation in Drosophila, Baz protein in the mesoderm is down-regulated from junctional sites in response to Snail (Sna) expression. This down-regulation leads to a specific decrease in adherens junctions without affecting other E-Cadherin pools. However, we further show that, interactions between Baz and junctions are not unidirectional. During apical constriction and the internalization of the mesoderm, down-regulation of Baz is transiently blocked as adherens junctions shift apically and are strengthened in response to tension generated by contractile actomyosin. When such junction remodeling is prevented by down-regulating myosin, Baz is lost prematurely in mesodermal epithelium. During such apical shifts, Baz is initially left behind as the junction shifts position, but then re-accumulates at the new location of the junctions. On the dorsal side of the embryo, a similar pattern of myosin activity appears to limit the basal shift in junctions normally driven by Baz that controls epithelium folding. Our results suggest a model where the sensitivity of Baz to Sna expression leads to the Sna-dependent junction disassembly required for a complete epithelium-mesenchymal transition. Meanwhile this loss of Baz-dependent junction maintenance is countered by the myosin-based mechanism which promotes an apical shift and strengthening of junctions accompanied by a transient re-positioning and maintenance of Baz proteins.


Subject(s)
Adherens Junctions/physiology , Drosophila Proteins/genetics , Drosophila melanogaster/embryology , Intracellular Signaling Peptides and Proteins/genetics , Membrane Proteins/genetics , Snail Family Transcription Factors/biosynthesis , Actomyosin/metabolism , Animals , Down-Regulation , Drosophila Proteins/biosynthesis , Epithelial-Mesenchymal Transition/physiology , Epithelium/metabolism , Female , Intracellular Signaling Peptides and Proteins/biosynthesis , Male , Mesoderm/physiology
17.
Proc Natl Acad Sci U S A ; 114(5): 1051-1056, 2017 01 31.
Article in English | MEDLINE | ID: mdl-28096360

ABSTRACT

Many models of morphogenesis are forced to assume specific mechanical properties of cells, because the actual mechanical properties of living tissues are largely unknown. Here, we measure the rheology of epithelial cells in the cellularizing Drosophila embryo by injecting magnetic particles and studying their response to external actuation. We establish that, on timescales relevant to epithelial morphogenesis, the cytoplasm is predominantly viscous, whereas the cellular cortex is elastic. The timescale of elastic stress relaxation has a lower bound of 4 min, which is comparable to the time required for internalization of the ventral furrow during gastrulation. The cytoplasm was measured to be ∼103-fold as viscous as water. We show that elasticity depends on the actin cytoskeleton and conclude by discussing how these results relate to existing mechanical models of morphogenesis.


Subject(s)
Drosophila melanogaster/embryology , Embryo, Nonmammalian/physiology , Epithelial Cells/physiology , Magnetite Nanoparticles , Magnets , Animals , Cytoplasm/drug effects , Cytoplasm/physiology , Cytoskeleton/physiology , Elasticity , Embryo, Nonmammalian/ultrastructure , Gastrulation/physiology , Giant Cells/physiology , Magnetics , Microinjections , Models, Biological , Morphogenesis , Rheology , Stress, Mechanical , Viscosity
18.
Elife ; 52016 11 23.
Article in English | MEDLINE | ID: mdl-27879204

ABSTRACT

During embryogenesis, the initial chromatin state is established during a period of rapid proliferative activity. We have measured with 3-min time resolution how heritable patterns of chromatin structure are initially established and maintained during the midblastula transition (MBT). We find that regions of accessibility are established sequentially, where enhancers are opened in advance of promoters and insulators. These open states are stably maintained in highly condensed mitotic chromatin to ensure faithful inheritance of prior accessibility status across cell divisions. The temporal progression of establishment is controlled by the biological timers that control the onset of the MBT. In general, acquisition of promoter accessibility is controlled by the biological timer that measures the nucleo-cytoplasmic (N:C) ratio, whereas timing of enhancer accessibility is regulated independently of the N:C ratio. These different timing classes each associate with binding sites for two transcription factors, GAGA-factor and Zelda, previously implicated in controlling chromatin accessibility at ZGA.


Subject(s)
Chromatin/chemistry , DNA-Binding Proteins/genetics , Drosophila Proteins/genetics , Drosophila melanogaster/genetics , Embryonic Development/genetics , Inheritance Patterns , Transcription Factors/genetics , Animals , Binding Sites , Biological Clocks/genetics , Cell Nucleus/metabolism , Chromatin/metabolism , Cytosol/metabolism , DNA-Binding Proteins/metabolism , Drosophila Proteins/metabolism , Drosophila melanogaster/growth & development , Drosophila melanogaster/metabolism , Embryo, Nonmammalian , Enhancer Elements, Genetic , Insulator Elements , Nuclear Proteins , Promoter Regions, Genetic , Protein Binding , Transcription Factors/metabolism
19.
Cell Rep ; 16(11): 2793-2801, 2016 09 13.
Article in English | MEDLINE | ID: mdl-27626650

ABSTRACT

The development of an embryo requires precise spatiotemporal regulation of cellular processes. During Drosophila gastrulation, a precise temporal pattern of cell division is encoded through transcriptional regulation of cdc25(string) in 25 distinct mitotic domains. Using a genetic screen, we demonstrate that the same transcription factors that regulate the spatial pattern of cdc25(string) transcription encode its temporal activation. We identify buttonhead and empty spiracles as the major activators of cdc25(string) expression in mitotic domain 2. The effect of these activators is balanced through repression by hairy, sloppy paired 1, and huckebein. Within the mitotic domain, temporal precision of mitosis is robust and unaffected by changing dosage of rate-limiting transcriptional factors. However, precision can be disrupted by altering the levels of the two activators or two repressors. We propose that the additive and balanced action of activators and repressors is a general strategy for precise temporal regulation of cellular transitions during development.


Subject(s)
Drosophila melanogaster/cytology , Drosophila melanogaster/embryology , Embryo, Nonmammalian/cytology , Mitosis/genetics , Transcription, Genetic , Animals , Base Pairing/genetics , Cell Lineage/genetics , Drosophila Proteins/genetics , Drosophila Proteins/metabolism , Drosophila melanogaster/genetics , Embryo, Nonmammalian/metabolism , Enhancer Elements, Genetic/genetics , Gene Dosage , Gene Expression Regulation, Developmental , Heterozygote , Mutation/genetics , Time Factors , Transcription Factors/metabolism
20.
Annu Rev Cell Dev Biol ; 32: 1-46, 2016 10 06.
Article in English | MEDLINE | ID: mdl-27501451

ABSTRACT

In large-scale mutagenesis screens performed in 1979-1980 at the EMBL in Heidelberg, we isolated mutations affecting the pattern or structure of the larval cuticle in Drosophila. The 600 mutants we characterized could be assigned to 120 genes and represent the majority of such genes in the genome. These mutants subsequently provided a rich resource for understanding many fundamental developmental processes, such as the transcriptional hierarchies controlling segmentation, the establishment of cell states by signaling pathways, and the differentiation of epithelial cells. Most of the Heidelberg genes are now molecularly known, and many of them are conserved in other animals, including humans. Although the screens were initially driven entirely by curiosity, the mutants now serve as models for many human diseases. In this review, we describe the rationale of the screening procedures and provide a classification of the genes on the basis of their initial phenotypes and the subsequent molecular analyses.


Subject(s)
Drosophila/genetics , Genetic Testing , Mutation/genetics , Animals , Genes, Insect , Mutagenesis/genetics , Signal Transduction/genetics
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