Developmental Regulation of the Cell Cycle in Drosophila

果蝇细胞周期的发育调控

基本信息

批准号：
7906559
负责人：
LAURA ANNE LEE
金额：
$ 6.26万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2009
资助国家：
美国
起止时间：
2009-09-01 至 2010-02-28
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7906559
关键词：
Address Animal Model Biochemical Biological Cell Cycle Cell Cycle Regulation Cell Nucleus Cellular biology Characteristics Chromosome Condensation Complex Cyclin B Cyclins DNA DNA biosynthesis Development Dimerization Drosophila genus Drosophila melanogaster Embryo Embryonic Development Ensure Enzymes Female Female sterility Genes Genetic Genome Homologous Gene In Vitro Methods Mitosis Mitotic Modeling Modification Mutation Pan Genus Phase Phenotype Phosphorus Phosphorylation Phosphorylation Site Phosphotransferases Physiological Plutonium Polyploidy Protein-Serine-Threonine Kinases Proteins RNA RNA Interference Regulation Role Serine Severities Staging Sterility Tacrolimus Binding Proteins Testing Ubiquitination anaphase-promoting complex base blastomere structure egg expression cloning gene conservation genome wide association study genome-wide human TYRP1 protein in vitro Assay in vivo multicatalytic endopeptidase complex mutant novel

项目摘要

DESCRIPTION (provided by applicant): The high degree of functional conservation of genes involved in the cell cycle combined with the superb cell biology and genetics of Drosophila make it an ideal model organism for studying cell cycle regulation. During early Drosophila embryogenesis, 3 genes are required for regulation of the streamlined "S-M" cycles characteristic of this developmental stage: pan gu (png), plutonium (plu), and giant nuclei (gnu). Females carrying mutations in any of these genes are sterile because their embryos fail to enter mitosis and over-replicate DNA to form "giant nuclei." PNG encodes a serine/threonine protein kinase, and plu and gnu encode small novel proteins, png, plu, and gnu interact genetically with cyclin B, and Cyclin B protein levels are decreased in these mutants. We recently demonstrated that PNG, PLU, and GNU interact to form an active kinase complex in vitro and that GNU activates PNG by inducing dimerization of PNG-PLU complexes. Additionally, we showed that GNU itself is a substrate for PNG kinase. In a genome-wide biochemical screen, several PNG kinase substrates with vertebrate homologs were recently identified, suggesting that the PNG kinase complex regulates the cell cycle by impinging on evolutionarily conserved components. We now propose to determine whether GNU activates PNG kinase via a dimerization mechanism in vivo and to test whether it is a substrate with potential function as an effector. We plan to determine whether the PNG kinase complex regulates activity of the Anaphase Promoting Complex so as to control Cyclin B levels and entry into mitosis. Finally, an evolutionarily conserved protein, Mat89Bb, has been identified as an in vitro PNG kinase substrate. RNA interference of Mat89Bb in Drosophila results in a phenotype reminiscent of that of the giant nuclei class of genes. We will test whether Mat89Bb is an in vitro substrate of PNG and will isolate Mat89Bb mutants to more precisely determine its role in cell cycle regulation.

描述（由申请人提供）：与细胞周期相关的基因的高度功能保护与果蝇的精湛细胞生物学和遗传学相结合，使其成为研究细胞周期调节的理想模型生物。在果蝇早期的胚胎发生过程中，需要3个基因来调节该发育阶段的简化“ S-M”周期特征：PAN GU（PNG），plu（PLU）和巨型核（GNU）。这些基因中携带突变的雌性都是无菌的，因为其胚胎未能进入有丝分裂和过度重新染色的DNA形成“巨型核”。 PNG编码丝氨酸/苏氨酸蛋白激酶，PLU和GNU编码小型新颖蛋白，PNG，PLU和GNU与细胞周期蛋白B遗传相互作用，细胞周期蛋白B蛋白水平在这些突变体中降低。我们最近证明了PNG，PLU和GNU相互作用以在体外形成活性激酶复合物，并且GNU通过诱导PNG-Plu复合物的二聚化来激活PNG。此外，我们表明GNU本身是PNG激酶的底物。在全基因组生化筛选中，最近确定了几种具有脊椎动物同源物的PNG激酶底物，这表明PNG激酶复合物通过撞击进化保守的成分来调节细胞周期。现在，我们建议确定GNU是否通过体内二聚化机制激活PNG激酶，并测试它是否是具有效应作为效应子的底物。我们计划确定PNG激酶复合物是否调节促进复合物的过期活性，从而控制细胞周期蛋白B水平并进入有丝分裂。最后，一种进化保守的蛋白MAT89BB已被确定为体外PNG激酶底物。 MAT89BB在果蝇中的RNA干扰会导致表型让人联想到巨型基因类别的基因。我们将测试MAT89BB是否是PNG的体外底物，并将分离MAT89BB突变体以更精确地确定其在细胞周期调节中的作用。