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1.
Cell ; 157(5): 1146-59, 2014 May 22.
Artigo em Inglês | MEDLINE | ID: mdl-24855950

RESUMO

E-cadherin is a major homophilic cell-cell adhesion molecule that inhibits motility of individual cells on matrix. However, its contribution to migration of cells through cell-rich tissues is less clear. We developed an in vivo sensor of mechanical tension across E-cadherin molecules, which we combined with cell-type-specific RNAi, photoactivatable Rac, and morphodynamic profiling, to interrogate how E-cadherin contributes to collective migration of cells between other cells. Using the Drosophila ovary as a model, we found that adhesion between border cells and their substrate, the nurse cells, functions in a positive feedback loop with Rac and actin assembly to stabilize forward-directed protrusion and directionally persistent movement. Adhesion between individual border cells communicates direction from the lead cell to the followers. Adhesion between motile cells and polar cells holds the cluster together and polarizes each individual cell. Thus, E-cadherin is an integral component of the guidance mechanisms that orchestrate collective chemotaxis in vivo.


Assuntos
Caderinas/metabolismo , Movimento Celular , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Animais , Fenômenos Biomecânicos , Adesão Celular , Quimiotaxia , Técnicas Citológicas , Drosophila melanogaster/metabolismo , Feminino , Dados de Sequência Molecular , Ovário/citologia , Proteínas rac de Ligação ao GTP/metabolismo
2.
PLoS Genet ; 16(12): e1009228, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33296356

RESUMO

Signal transduction pathways are intricately fine-tuned to accomplish diverse biological processes. An example is the conserved Ras/mitogen-activated-protein-kinase (MAPK) pathway, which exhibits context-dependent signaling output dynamics and regulation. Here, by altering codon usage as a novel platform to control signaling output, we screened the Drosophila genome for modifiers specific to either weak or strong Ras-driven eye phenotypes. Our screen enriched for regions of the genome not previously connected with Ras phenotypic modification. We mapped the underlying gene from one modifier to the ribosomal gene RpS21. In multiple contexts, we show that RpS21 preferentially influences weak Ras/MAPK signaling outputs. These data show that codon usage manipulation can identify new, output-specific signaling regulators, and identify RpS21 as an in vivo Ras/MAPK phenotypic regulator.


Assuntos
Uso do Códon , Proteínas de Drosophila/genética , Genes Modificadores , Proteínas Quinases Ativadas por Mitógeno/genética , Proteínas ras/genética , Animais , Proteínas de Drosophila/metabolismo , Drosophila melanogaster , Sistema de Sinalização das MAP Quinases , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Proteínas ras/metabolismo
3.
Development ; 144(22): 4091-4102, 2017 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-28947534

RESUMO

The molecular identities and regulation of cells at interorgan boundaries are often unclear, despite the increasingly appreciated role of organ boundaries in disease. Using Drosophila as a model, we here show that a specific population of adult midgut organ-boundary intestinal stem cells (OB-ISCs) is regulated by the neighboring hindgut, a developmentally distinct organ. This distinct OB-ISC control occurs through proximity to a specialized transition zone between the endodermal midgut and ectodermal hindgut that shares molecular signatures of both organs, which we term the hybrid zone (HZ). During homeostasis, proximity to the HZ restrains OB-ISC proliferation. However, injury to the adult HZ/hindgut drives upregulation of unpaired-3 cytokine, which signals through a Signal transducer and activator of transcription (STAT) protein to promote cell division only in OB-ISCs. If HZ disruption is severe, hyperplastic OB-ISCs expand across the interorgan boundary. Our data suggest that interorgan signaling plays an important role in controlling OB-ISCs in homeostasis and injury repair, which is likely to be crucial in prevention of disease.


Assuntos
Padronização Corporal , Drosophila melanogaster/citologia , Drosophila melanogaster/embriologia , Intestinos/citologia , Especificidade de Órgãos , Células-Tronco/citologia , Animais , Carcinogênese/patologia , Ciclo Celular , Proliferação de Células , Proteínas de Drosophila/metabolismo , Hiperplasia , Intestinos/crescimento & desenvolvimento , Janus Quinases/metabolismo , Larva/fisiologia , Modelos Biológicos , Fatores de Transcrição STAT/metabolismo , Transdução de Sinais
4.
Nat Commun ; 15(1): 5270, 2024 Jun 20.
Artigo em Inglês | MEDLINE | ID: mdl-38902233

RESUMO

Regulation of codon optimality is an increasingly appreciated layer of cell- and tissue-specific protein expression control. Here, we use codon-modified reporters to show that differentiation of Drosophila neural stem cells into neurons enables protein expression from rare-codon-enriched genes. From a candidate screen, we identify the cytoplasmic polyadenylation element binding (CPEB) protein Orb2 as a positive regulator of rare-codon-dependent mRNA stability in neurons. Using RNA sequencing, we reveal that Orb2-upregulated mRNAs in the brain with abundant Orb2 binding sites have a rare-codon bias. From these Orb2-regulated mRNAs, we demonstrate that rare-codon enrichment is important for mRNA stability and social behavior function of the metabotropic glutamate receptor (mGluR). Our findings reveal a molecular mechanism by which neural stem cell differentiation shifts genetic code regulation to enable critical mRNA stability and protein expression.


Assuntos
Diferenciação Celular , Proteínas de Drosophila , Células-Tronco Neurais , Neurônios , Estabilidade de RNA , RNA Mensageiro , Animais , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/genética , Neurônios/metabolismo , Neurônios/citologia , RNA Mensageiro/metabolismo , RNA Mensageiro/genética , Diferenciação Celular/genética , Células-Tronco Neurais/metabolismo , Células-Tronco Neurais/citologia , Códon/genética , Drosophila melanogaster/genética , Drosophila melanogaster/citologia , Drosophila melanogaster/metabolismo , Receptores de Glutamato Metabotrópico/metabolismo , Receptores de Glutamato Metabotrópico/genética , Fatores de Poliadenilação e Clivagem de mRNA/metabolismo , Fatores de Poliadenilação e Clivagem de mRNA/genética , Drosophila/genética , Drosophila/metabolismo , Encéfalo/metabolismo , Encéfalo/citologia , Fatores de Transcrição
5.
bioRxiv ; 2023 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-37546801

RESUMO

Regulation of codon optimality is an increasingly appreciated layer of cell- and tissue-specific protein expression control. Here, we use codon-modified reporters to show that differentiation of Drosophila neural stem cells into neurons enables protein expression from rare-codon-enriched genes. From a candidate screen, we identify the cytoplasmic polyadenylation element binding (CPEB) protein Orb2 as a positive regulator of rare-codon-dependent expression in neurons. Using RNA sequencing, we reveal that Orb2-upregulated mRNAs in the brain with abundant Orb2 binding sites have a rare-codon bias. From these Orb2-regulated mRNAs, we demonstrate that rare-codon enrichment is important for expression control and social behavior function of the metabotropic glutamate receptor (mGluR). Our findings reveal a molecular mechanism by which neural stem cell differentiation shifts genetic code regulation to enable critical mRNA and protein expression.

6.
Elife ; 112022 05 06.
Artigo em Inglês | MEDLINE | ID: mdl-35522036

RESUMO

Codon usage bias has long been appreciated to influence protein production. Yet, relatively few studies have analyzed the impacts of codon usage on tissue-specific mRNA and protein expression. Here, we use codon-modified reporters to perform an organism-wide screen in Drosophila melanogaster for distinct tissue responses to codon usage bias. These reporters reveal a cliff-like decline of protein expression near the limit of rare codon usage in endogenously expressed Drosophila genes. Near the edge of this limit, however, we find the testis and brain are uniquely capable of expressing rare codon-enriched reporters. We define a new metric of tissue-specific codon usage, the tissue-apparent Codon Adaptation Index (taCAI), to reveal a conserved enrichment for rare codon usage in the endogenously expressed genes of both Drosophila and human testis. We further demonstrate a role for rare codons in an evolutionarily young testis-specific gene, RpL10Aa. Optimizing RpL10Aa codons disrupts female fertility. Our work highlights distinct responses to rarely used codons in select tissues, revealing a critical role for codon bias in tissue biology.


Assuntos
Drosophila melanogaster , Drosophila , Animais , Códon/genética , Uso do Códon , Drosophila/genética , Drosophila melanogaster/genética , Feminino , Humanos , Masculino , Testículo
7.
Dev Cell ; 56(14): 2059-2072.e3, 2021 07 26.
Artigo em Inglês | MEDLINE | ID: mdl-34019841

RESUMO

Individual organ development must be temporally coordinated with development of the rest of the organism. As a result, cell division cycles in a developing organ occur on a relatively fixed timescale. Despite this, many developing organs can regenerate cells lost to injury. How organs regenerate within the time constraints of organism development remains unclear. Here, we show that the developing Drosophila hindgut regenerates by accelerating the mitotic cell cycle. This process is achieved by decreasing G1 length and requires the JAK/STAT ligand unpaired-3. Mitotic capacity is then terminated by the steroid hormone ecdysone receptor and the Sox transcription factor Dichaete. These two factors converge on regulation of a hindgut-specific enhancer of fizzy-related, a negative regulator of mitotic cyclins. Our findings reveal how the cell-cycle machinery and cytokine signaling can be adapted to accomplish developmental organ regeneration.


Assuntos
Proteínas de Drosophila/metabolismo , Drosophila melanogaster/crescimento & desenvolvimento , Fase G1 , Trato Gastrointestinal/crescimento & desenvolvimento , Regulação da Expressão Gênica no Desenvolvimento , Organogênese , Regeneração , Animais , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/metabolismo , Feminino , Trato Gastrointestinal/lesões , Trato Gastrointestinal/metabolismo , Janus Quinases/genética , Janus Quinases/metabolismo , Masculino , Mitose , Fatores de Transcrição SOX/genética , Fatores de Transcrição SOX/metabolismo , Fatores de Transcrição STAT/genética , Fatores de Transcrição STAT/metabolismo , Fatores de Transcrição/genética , Fatores de Transcrição/metabolismo
8.
Genetics ; 214(2): 235-264, 2020 02.
Artigo em Inglês | MEDLINE | ID: mdl-32029579

RESUMO

The insect excretory system contains two organ systems acting in concert: the Malpighian tubules and the hindgut perform essential roles in excretion and ionic and osmotic homeostasis. For over 350 years, these two organs have fascinated biologists as a model of organ structure and function. As part of a recent surge in interest, research on the Malpighian tubules and hindgut of Drosophila have uncovered important paradigms of organ physiology and development. Further, many human disease processes can be modeled in these organs. Here, focusing on discoveries in the past 10 years, we provide an overview of the anatomy and physiology of the Drosophila excretory system. We describe the major developmental events that build these organs during embryogenesis, remodel them during metamorphosis, and repair them following injury. Finally, we highlight the use of the Malpighian tubules and hindgut as accessible models of human disease biology. The Malpighian tubule is a particularly excellent model to study rapid fluid transport, neuroendocrine control of renal function, and modeling of numerous human renal conditions such as kidney stones, while the hindgut provides an outstanding model for processes such as the role of cell chirality in development, nonstem cell-based injury repair, cancer-promoting processes, and communication between the intestine and nervous system.


Assuntos
Eliminação Intestinal/fisiologia , Túbulos de Malpighi/metabolismo , Túbulos de Malpighi/fisiologia , Animais , Modelos Animais de Doenças , Drosophila/metabolismo , Proteínas de Drosophila/metabolismo , Desenvolvimento Embrionário , Endoderma , Homeostase , Mucosa Intestinal/metabolismo , Intestinos/fisiologia
9.
Elife ; 72018 08 17.
Artigo em Inglês | MEDLINE | ID: mdl-30117808

RESUMO

Ploidy-increasing cell cycles drive tissue growth in many developing organs. Such cycles, including endocycles, are increasingly appreciated to drive tissue growth following injury or activated growth signaling in mature organs. In these organs, the regulation and distinct roles of different cell cycles remains unclear. Here, we uncover a programmed switch between cell cycles in the Drosophila hindgut pylorus. Using an acute injury model, we identify mitosis as the response in larval pyloric cells, whereas endocycles occur in adult pyloric cells. By developing a novel genetic method, DEMISE (Dual-Expression-Method-for-Induced-Site-specific-Eradication), we show the cell cycle regulator Fizzy-related dictates the decision between mitosis and endocycles. After injury, both cycles accurately restore tissue mass and genome content. However, in response to sustained growth signaling, only endocycles preserve epithelial architecture. Our data reveal distinct cell cycle programming in response to similar stimuli in mature vs. developmental states and reveal a tissue-protective role of endocycles.


Assuntos
Proteínas Cdh1/genética , Ciclo Celular/genética , Sistema Digestório/crescimento & desenvolvimento , Proteínas de Drosophila/genética , Mucosa Gástrica/crescimento & desenvolvimento , Animais , Proteínas de Ciclo Celular/genética , Sistema Digestório/citologia , Drosophila melanogaster/genética , Drosophila melanogaster/crescimento & desenvolvimento , Mucosa Gástrica/citologia , Regulação da Expressão Gênica no Desenvolvimento , Larva/genética , Larva/crescimento & desenvolvimento , Mitose/genética , Transdução de Sinais/genética
10.
Mol Biol Cell ; 22(14): 2491-508, 2011 Jul 15.
Artigo em Inglês | MEDLINE | ID: mdl-21613546

RESUMO

Integrating individual cell movements to create tissue-level shape change is essential to building an animal. We explored mechanisms of adherens junction (AJ):cytoskeleton linkage and roles of the linkage regulator Canoe/afadin during Drosophila germband extension (GBE), a convergent-extension process elongating the body axis. We found surprising parallels between GBE and a quite different morphogenetic movement, mesoderm apical constriction. Germband cells have an apical actomyosin network undergoing cyclical contractions. These coincide with a novel cell shape change--cell extension along the anterior-posterior (AP) axis. In Canoe's absence, GBE is disrupted. The apical actomyosin network detaches from AJs at AP cell borders, reducing coordination of actomyosin contractility and cell shape change. Normal GBE requires planar polarization of AJs and the cytoskeleton. Canoe loss subtly enhances AJ planar polarity and dramatically increases planar polarity of the apical polarity proteins Bazooka/Par3 and atypical protein kinase C. Changes in Bazooka localization parallel retraction of the actomyosin network. Globally reducing AJ function does not mimic Canoe loss, but many effects are replicated by global actin disruption. Strong dose-sensitive genetic interactions between canoe and bazooka are consistent with them affecting a common process. We propose a model in which an actomyosin network linked at AP AJs by Canoe and coupled to apical polarity proteins regulates convergent extension.


Assuntos
Actomiosina/metabolismo , Junções Aderentes/metabolismo , Proteínas de Drosophila/genética , Drosophila melanogaster/genética , Drosophila melanogaster/fisiologia , Actomiosina/genética , Actomiosina/fisiologia , Junções Aderentes/genética , Junções Aderentes/fisiologia , Animais , Movimento Celular/genética , Movimento Celular/fisiologia , Polaridade Celular/genética , Polaridade Celular/fisiologia , Forma Celular/genética , Forma Celular/fisiologia , Citoesqueleto/genética , Citoesqueleto/fisiologia , Proteínas de Drosophila/metabolismo , Proteínas de Drosophila/fisiologia , Gastrulação/genética , Mesoderma/crescimento & desenvolvimento , Morfogênese/genética , Morfogênese/fisiologia , Mutação
11.
PLoS One ; 4(10): e7634, 2009 Oct 28.
Artigo em Inglês | MEDLINE | ID: mdl-19862327

RESUMO

BACKGROUND: Tissue morphogenesis and organogenesis require that cells retain stable cell-cell adhesion while changing shape and moving. One mechanism to accommodate this plasticity in cell adhesion involves regulated trafficking of junctional proteins. METHODOLOGY/PRINCIPAL FINDINGS: Here we explored trafficking of junctional proteins in two well-characterized model epithelia, the Drosophila embryonic ectoderm and amnioserosa. We find that DE-cadherin, the transmembrane protein of adherens junctions, is actively trafficked through putative vesicles, and appears to travel through both Rab5-positive and Rab11-positive structures. We manipulated the functions of Rab11 and Rab5 to examine the effects on junctional stability and morphogenesis. Reducing Rab11 function, either using a dominant negative construct or loss of function alleles, disrupts integrity of the ectoderm and leads to loss of adherens junctions. Strikingly, the apical junctional regulator Crumbs is lost before AJs are destabilized, while the basolateral protein Dlg remains cortical. Altering Rab5 function had less dramatic effects, not disrupting adherens junction integrity but affecting dorsal closure. CONCLUSIONS/SIGNIFICANCE: We contrast our results with what others saw when disrupting other trafficking regulators, and when disrupting Rab function in other tissues; together these data suggest distinct mechanisms regulate junctional stability and plasticity in different tissues.


Assuntos
Junções Aderentes/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/embriologia , Drosophila melanogaster/genética , Ectoderma/metabolismo , Regulação da Expressão Gênica no Desenvolvimento , Proteínas de Membrana/metabolismo , Proteínas rab de Ligação ao GTP/metabolismo , Alelos , Animais , Animais Geneticamente Modificados , Adesão Celular , Cruzamentos Genéticos , Endocitose , Genes Dominantes , Genes de Insetos , Microscopia de Fluorescência/métodos , Mutação
12.
Curr Top Dev Biol ; 89: 55-85, 2009.
Artigo em Inglês | MEDLINE | ID: mdl-19737642

RESUMO

One key challenge for cell and developmental biologists is to determine how the cytoskeletal toolkit is used to build embryonic tissues and organs. Here, we review recent progress in meeting this challenge, focusing on epithelial morphogenesis in the Drosophila embryo as a model. We outline how actin and microtubule networks are regulated by embryonic patterning systems, and how they affect cell shape, cell behavior, and cell-cell interactions to shape epithelial structures. We focus on the formation of the first epithelium at cellularization, the assembly of junctions, apical constriction of cells in the ventral furrow, cell intercalation in the germband, and epithelial sheet migration during dorsal closure. These events provide models for uncovering the cell biological basis of morphogenesis.


Assuntos
Padronização Corporal , Citoesqueleto/metabolismo , Drosophila melanogaster/embriologia , Embrião não Mamífero/embriologia , Animais , Epitélio/embriologia
13.
J Cell Biol ; 186(1): 57-73, 2009 Jul 13.
Artigo em Inglês | MEDLINE | ID: mdl-19596848

RESUMO

Cadherin-based adherens junctions (AJs) mediate cell adhesion and regulate cell shape change. The nectin-afadin complex also localizes to AJs and links to the cytoskeleton. Mammalian afadin has been suggested to be essential for adhesion and polarity establishment, but its mechanism of action is unclear. In contrast, Drosophila melanogaster's afadin homologue Canoe (Cno) has suggested roles in signal transduction during morphogenesis. We completely removed Cno from embryos, testing these hypotheses. Surprisingly, Cno is not essential for AJ assembly or for AJ maintenance in many tissues. However, morphogenesis is impaired from the start. Apical constriction of mesodermal cells initiates but is not completed. The actomyosin cytoskeleton disconnects from AJs, uncoupling actomyosin constriction and cell shape change. Cno has multiple direct interactions with AJ proteins, but is not a core part of the cadherin-catenin complex. Instead, Cno localizes to AJs by a Rap1- and actin-dependent mechanism. These data suggest that Cno regulates linkage between AJs and the actin cytoskeleton during morphogenesis.


Assuntos
Actinas/metabolismo , Junções Aderentes/metabolismo , Polaridade Celular , Citoesqueleto/metabolismo , Proteínas de Drosophila/metabolismo , Drosophila melanogaster/citologia , Proteínas dos Microfilamentos/química , Actomiosina/metabolismo , Animais , Caderinas/metabolismo , Extensões da Superfície Celular/metabolismo , Proteínas de Drosophila/deficiência , Drosophila melanogaster/embriologia , Drosophila melanogaster/metabolismo , Mesoderma/metabolismo , Morfogênese , Mutação/genética , Especificidade de Órgãos , Ligação Proteica , Transporte Proteico , Coelhos , Homologia de Sequência de Aminoácidos , alfa Catenina/metabolismo , Proteínas rap1 de Ligação ao GTP/metabolismo
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