Regulation and function of PIASy mediated mitotic SUMOylation in vertebrates

PIASy 介导的脊椎动物有丝分裂 SUMO 化的调节和功能

基本信息

批准号：
8071587
负责人：
Yoshiaki Azuma
金额：
$ 36.36万
依托单位：
UNIVERSITY OF KANSAS LAWRENCE
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-07-01 至 2013-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8071587
关键词：
Anaphase Binding Binding Proteins Biochemical Biological Assay Biological Models Cell Cycle Cell Cycle Regulation Cell division Cell physiology Cells Centromere Chromatin Chromosome Segregation Chromosome Structures Chromosome abnormality Chromosomes Complex Defect Development Drosophila genus Event Failure Family Genetic Genomics Hela Cells In Vitro Indium Investigation Ligase Mediating Mitosis Mitotic Mitotic Chromosome Modification Molecular Mutation Nuclear Nuclear Structure Pathway interactions Phosphorylation Physiological Play Poly Adenosine Diphosphate Ribose Post-Translational Protein Processing Process Prometaphase Protein Family Proteins RNA Interference Regulation Research Resistance Role Scaffolding Protein Sodium Chloride Sumoylation Pathway System Topoisomerase II Transcriptional Regulation Ubiquitin Vertebrates Xenopus Yeasts aurora B kinase daughter cell egg in vivo member mutant neutralizing antibody novel protein inhibitors of activated STAT research study segregation tumor progression ubiquitin-protein ligase

项目摘要

DESCRIPTION (provided by applicant): During cell division, the full set of chromosomes needs to separate accurately at anaphase to maintain complete genomic information in each daughter cell. Failure of proper chromosome segregation at anaphase leads to biased genomic information in daughter cells, which causes developmental defect and contributes to tumor progression. Precise structural organization of mitotic chromosomes is a key element in maintaining integrity of chromosome segregation, which is regulated by multiple posttranslational protein modification systems. Both genetic and biochemical studies indicate that the posttranslational protein modification by Small Ubiquitin-like Modifier (SUMO) modification pathway has an important role in normal progression of mitosis. I have found that mitotic specific SUMO-2 modification has a crucial role for completion of faithful chromosome segregation in anaphase in Xenopus egg extract assay system. This mitotic SUMO-2 modification specifically requires the activity of the PIASy protein that is a member of conserved PIAS family of E3. PIASy mediates SUMO-2 modification on multiple mitotic chromosomal proteins. Inhibition of SUMOylation in mitosis by either addition of mutant form of Ubc9, SUMO E2, or anti-PIASy neutralizing antibodies causes compromised chromosome segregation at anaphase. The major PIASy-mediated SUMO-2 substrate during mitosis is DMA topoisomerase II (Topoll), which is known to have an essential function in organizing mitotic chromosomes. Inhibition of SUMOylation alters the chromosomal association status of Topoll. I hypothesize that the PIASy mediated mitotic SUMOylation has an essential function in regulating the organization of mitotic chromosomes via its substrates, which plays a crucial role in chromosome segregation at anaphase. I propose to investigate a regulatory mechanism of PIASy specific SUMOylation pathway (Aim1) and a consequence of mitotic SUMOylation with determining the function of Topoll SUMOylation (Aim2) and identifying novel PIASy substrates (Aim3) by using Xenopus egg extracts as a model system. This investigation will demonstrate the function of SUMO-2 modification in organization of mitotic chromosomes in vertebrates. Determination of regulatory mechanisms of PIASy mediated SUMOylation will provide an invaluable information for understanding SUMOylation pathway that has an impact on diverse cellular physiological functions.

描述（由申请人提供）：在细胞分裂过程中，全套染色体需要在后期准确分离，以维持每个子细胞中完整的基因组信息。后期染色体分离失败会导致子细胞中的基因组信息出现偏差，从而导致发育缺陷并导致肿瘤进展。有丝分裂染色体的精确结构组织是维持染色体分离完整性的关键要素，其受到多个翻译后蛋白质修饰系统的调节。遗传和生化研究表明，小泛素样修饰剂（SUMO）修饰途径的翻译后蛋白修饰在有丝分裂的正常进展中具有重要作用。我发现有丝分裂特异性 SUMO-2 修饰对于爪蟾卵提取物测定系统中后期染色体忠实分离的完成具有至关重要的作用。这种有丝分裂 SUMO-2 修饰特别需要 PIASy 蛋白的活性，PIASy 蛋白是 E3 保守 PIAS 家族的成员。 PIASy 介导 SUMO-2 对多种有丝分裂染色体蛋白的修饰。通过添加 Ubc9、SUMO E2 突变体或抗 PIASy 中和抗体抑制有丝分裂中的 SUMO 化，导致后期染色体分离受损。有丝分裂过程中 PIASy 介导的 SUMO-2 的主要底物是 DMA 拓扑异构酶 II (Topoll)，已知它在组织有丝分裂染色体中具有重要功能。 SUMO 化的抑制会改变 Topoll 的染色体关联状态。我假设 PIASy 介导的有丝分裂 SUMO 化在通过其底物调节有丝分裂染色体的组织方面具有重要功能，这在后期染色体分离中起着至关重要的作用。我建议通过使用非洲爪蟾卵提取物作为模型系统来确定 Topoll SUMOylation 的功能 (Aim2) 并鉴定新型 PIASy 底物 (Aim3)，从而研究 PIASy 特异性 SUMOylation 途径 (Aim1) 的调节机制和有丝分裂 SUMOylation 的结果。这项研究将证明 SUMO-2 修饰在脊椎动物有丝分裂染色体组织中的功能。确定 PIASy 介导的 SUMO 化的调节机制将为了解影响多种细胞生理功能的 SUMO 化途径提供宝贵的信息。