Cell Cycle Regulation of Membrane Trafficking

膜运输的细胞周期调控

基本信息

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
项目类别：
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

项目摘要

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.

项目概要细胞分裂需要一系列仔细协调的基本事件，这些事件必须受到精确调控。细胞分裂的一个核心特征是将染色体准确地分离成子细胞。其他细胞的区室还必须小心地包装成子细胞，并与染色体分离。膜运输途径对于胞质分裂的完成至关重要细胞分裂结束。细胞在胞质分裂过程中如何控制膜运输尚不清楚。大的蛋白酶分离酶由于其在粘连蛋白切割中的作用而成为染色体分离的核心角色，允许染色体在后期开始时分离。染色体分离后，分离酶促进后期的几个事件。该提案旨在了解分离酶在胞吐作用中的新作用胞质分裂所需的 RAB-11 囊泡。分离酶还调节大皮质颗粒的胞吐作用在减数分裂 I 的后期阻止多精受精，这是分析这一调控途径的理想背景。我们将使用生化和遗传学方法来识别分离酶的底物或结合伴侣囊泡来定义其促进胞吐作用的机制。 separase的动态定位是在细胞分裂过程中受到调节，分离酶仅在后期定位于囊泡。我们将调查如何染色体分离调节因子控制分离酶活性和囊泡定位。过度表达不可降解的 securin 将用于确定这种抑制性伴侣如何控制胞吐功能的分离。 PPH-5 磷酸酶及其激活剂 HSP-90 的突变被确定为抑制因子分离酶突变体的胚胎致死率。将绘制分离酶磷酸化位点并进行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 将进行测试以确定它是否直接在体外使分离酶去磷酸化。 PPH-5 和 HSP-90 的功能将被表征为确定它们是否在减数分裂期间直接调节分离酶。这些研究将使用遗传易驯化的秀丽隐杆线虫胚胎进行。这项工作将为细胞如何在胞质分裂过程中协调染色体分离和胞吐作用的基本过程，即与了解正常发育和不孕症和癌症等疾病相关。

项目成果

期刊论文数量（13）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

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

BUB-1 靶向 PP2A:B56，以调节线虫卵母细胞减数分裂 I 期间的染色体聚集。

DOI：
发表时间：
2020-12-23
期刊：
影响因子：
7.7
作者：
Bel Borja, Laura;Soubigou, Flavie;Taylor, Samuel J P;Fraguas Bringas, Conchita;Budrewicz, Jacqueline;Lara;Sorensen Turpin, Christopher G;Bembenek, Joshua N;Cheerambathur, Dhanya K;Pelisch, Federico
通讯作者：
Pelisch, Federico

Meeting report - Cellular dynamics: membrane-cytoskeleton interface.

会议报告 - 细胞动力学：膜-细胞骨架界面。

DOI：
发表时间：
2017-09-01
期刊：
影响因子：
4
作者：
Bembenek, Joshua N;Meshik, Xenia;Tsarouhas, Vasilios
通讯作者：
Tsarouhas, Vasilios

Investigating the Roles of Master Cell Cycle Regulators during Cytokinesis and Embryonic Development in Caenorhabditis elegans

研究主细胞周期调节因子在秀丽隐杆线虫细胞分裂和胚胎发育过程中的作用

DOI：
发表时间：
2024-09-14
期刊：
影响因子：
0
作者：
X. Bai
通讯作者：
X. Bai

Aurora B functions at the apical surface after specialized cytokinesis during morphogenesis in C. elegans.

在线虫形态发生过程中，Aurora B 在专门的胞质分裂后在顶端表面发挥作用。

DOI：
发表时间：
2020-01-08
期刊：
影响因子：
0
作者：
Bai, Xiaofei;Melesse, Michael;Sorensen Turpin, Christopher G;Sloan, Dillon E;Chen, Chin;Wang, Wen;Lee, Po;Simmons, James R;Nebenfuehr, Benjamin;Mitchell, Diana;Klebanow, Lindsey R;Mattson, Nicholas;Betzig, Eric;Chen, Bi;Che
通讯作者：
Che

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

CAV-1::GFP 在秀丽隐杆线虫中的内源表达和定位。