Cell Cycle Regulation of Membrane Trafficking

膜运输的细胞周期调控

• 批准号: 10640131
• 负责人: Joshua Nathaniel Bembenek
• 金额: $ 29.61万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2015
• 资助国家: 美国
• 起止时间: 2015-03-15 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10640131
• 关键词: Address; Affect; Alleles; Anaphase; Binding; Biochemical; Caenorhabditis elegans; Cell Compartmentation; Cell Cycle; Cell Cycle Regulation; Cell division; Cells; Cellular biology; Chromosome Segregation; Chromosomes; Complex; Cytokinesis; Cytoplasmic Granules; Data; Development; Disease; Disparate; Embryo; Enzymes; Event; Exocytosis; Fertilization; Funding; Genetic; Genome; Goals; Heat-Shock Proteins 90; In Vitro; Infertility; Inherited; Knowledge; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Meiosis; Membrane; Mitotic; Molecular Chaperones; Mutation; Oocytes; PTTG1 gene; Pathway interactions; Peptide Hydrolases; Phosphoric Monoester Hydrolases; Phosphorylation; Phosphorylation Site; Process; Protein Dephosphorylation; Regulation; Regulation of Exocytosis; Regulatory Pathway; Role; Series; Sister Chromatid; Testing; Vesicle; Work; anaphase-promoting complex; cohesin; daughter cell; genetic approach; insight; mutant; novel; overexpression; prevent; segregation; separase; tool; trafficking

Project Summary Cell division requires a carefully coordinated series of essential events that must be precisely regulated. A central feature of cell division is the accurate segregation of chromosomes into daughter cells. Other compartments of the cell must also be carefully packaged into daughter cells, and coordinated with chromosome segregation. Membrane trafficking pathways are essential for the completion of cytokinesis at the end of cell division. How cells control membrane trafficking during cytokinesis is not well understood. The large protease separase is a central player in chromosome segregation due to its role in cohesin cleavage, which allows chromosome separation at the onset of anaphase. After chromosome segregation, separase promotes several events during anaphase. This proposal aims to understand a novel role of separase in the exocytosis of RAB-11 vesicles required for cytokinesis. Separase also regulates exocytosis of large cortical granules during anaphase of meiosis I to block polyspermy, which is an ideal context to analyze this regulatory pathway. We will use biochemical and genetic approaches to identify substrates or binding partners of separase on vesicles to define the mechanism by which it promotes exocytosis. The dynamic localization of separase is regulated during cell division and separase only localizes to vesicles during anaphase. We will investigate how chromosome segregation regulators control separase activity and localization to vesicles. Overexpression of non-degradable securin will be used to determine how this inhibitory chaperone controls the exocytic function of separase. Mutations of the PPH-5 phosphatase and its activator HSP-90 were identified as suppressors of embryo lethality of separase mutants. Separase phosphorylation sites will be mapped and phosphorylation mutants will be studied to determine how they affect separase function. PPH-5 will be tested to determine if it directly dephosphorylates separase in vitro. The functions of PPH-5 and HSP-90 will be characterized to determine whether they directly regulate separase during the meiotic divisions. These studies will be performed using the genetically tractable C. elegans embryo. This work will provide new insight into how cells coordinate the essential process of chromosome segregation with exocytosis during cytokinesis, which is relevant to understanding normal development and diseases such as infertility and cancer.

项目摘要 细胞分裂需要精心协调的一系列基本事件，必须精确调节。 细胞分裂的一个主要特征是将染色体的准确分离为子细胞。其他 细胞的隔室也必须小心地包装到子细胞中，并与 染色体分离。膜运输途径对于完成细胞因子至关重要 细胞分裂的末端。细胞如何控制细胞因子期间的膜运输尚不清楚。大 蛋白酶分离酶是染色体隔离的核心参与者，因为它在粘蛋白裂解中的作用 允许在后期开始时染色体分离。染色体分离后，分离酶促进 后期期间的几个事件。该建议旨在了解分离酶在胞吐作用中的新作用 rab-11囊泡的细胞因子。分离酶还调节大型皮质颗粒的胞吐作用 在减数分裂后期，I阻断多植物，这是分析该调节途径的理想背景。 我们将使用生化和遗传方法来鉴定分离酶的底物或结合伴侣 囊泡定义促进胞吐作用的机制。分离酶的动态定位是 在细胞分裂期间调节，分离酶仅在后期期间定位于囊泡。我们将调查如何 染色体分离调节剂控制分离酶的活性和囊泡的定位。过表达 不良券将用于确定这种抑制性伴侣如何控制外囊肿功能 分离酶。 PPH-5磷酸酶及其活化剂HSP-90的突变被确定为抑制剂 分离酶突变体的胚胎致死性。分离酶磷酸化位点将被映射并磷酸化 将研究突变体以确定它们如何影响分离酶功能。 PPH-5将测试以确定是否 直接在体外脱磷酸化酶。 PPH-5和HSP-90的功能将被表征为 确定它们是否在减数分裂分裂期间直接调节分离酶。这些研究将是 使用遗传性的秀丽隐杆线虫胚胎进行。这项工作将为细胞如何提供新的见解 在细胞因子期间，协调染色体分离的基本过程 与了解正常发育和疾病（例如不育和癌症）有关。

项目成果

PLK1- and PLK4-Mediated Asymmetric Mitotic Centrosome Size and Positioning in the Early Zebrafish Embryo.

• DOI: 10.1016/j.cub.2020.08.074
• 发表时间: 2020-11-16
• 期刊: Current biology : CB
• 作者: Rathbun LI;Aljiboury AA;Bai X;Hall NA;Manikas J;Amack JD;Bembenek JN;Hehnly H
• 通讯作者: Hehnly H

Endogenous expression and localization of CAV-1::GFP in C. elegans.

• DOI: 10.17912/micropub.biology.000311
• 发表时间: 2020-09-21
• 期刊: microPublication biology
• 作者: Sloan DE;Bembenek JN
• 通讯作者: Bembenek JN

Cracking the eggshell: A novel link to intracellular signaling.

打破蛋壳：细胞内信号传导的新联系。

• DOI: 10.1016/j.ydbio.2019.05.014
• 发表时间: 2019
• 期刊: Developmental biology
• 影响因子: 2.7
• 作者: Melesse,Michael;Bembenek,JoshuaN
• 通讯作者: Bembenek,JoshuaN

Genetic Identification of Separase Regulators in Caenorhabditis elegans.

秀丽隐杆线虫分离酶调节剂的遗传鉴定。

• DOI: 10.1534/g3.117.300298
• 发表时间: 2018
• 期刊: G3 (Bethesda, Md.)
• 作者: Melesse,Michael;Sloan,DillonE;Benthal,JosephT;Caylor,Quincey;Gosine,Krishen;Bai,Xiaofei;Bembenek,JoshuaN
• 通讯作者: Bembenek,JoshuaN

BUB-1 targets PP2A:B56 to regulate chromosome congression during meiosis I in C. elegans oocytes.

• DOI: 10.7554/elife.65307
• 发表时间: 2020-12-23
• 期刊: eLife
• 影响因子: 7.7
• 作者: Bel Borja L;Soubigou F;Taylor SJP;Fraguas Bringas C;Budrewicz J;Lara-Gonzalez P;Sorensen Turpin CG;Bembenek JN;Cheerambathur DK;Pelisch F
• 通讯作者: Pelisch F