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）：在细胞分裂期间，完整的染色体需要在后期准确分离，以维持每个子细胞中的完整基因组信息。适当的染色体分离在后期的失败会导致子细胞中的基因组信息有偏见，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隔离完整性的关键要素，该要素受多种翻译后蛋白质修饰系统调节。遗传学和生化研究都表明，小型泛素样修饰剂（SUMO）修饰途径的翻译后蛋白质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修饰对于在Xenopus Egg提取物测定系统中的后期完成忠实的染色体隔离至关重要。这种有丝分裂的SUMO-2修饰特异性要求具有E3的保守性PIA家族的纯化蛋白的活性。 Piasy介导了多丝分裂染色体蛋白上的SUMO-2修饰。通过添加UBC9，SUMO E2的突变形式或抗pia抗中和抗体会导致后期的染色体隔离导致抑制有丝分裂的Sumoylation。有丝分裂过程中的主要旋律介导的SUMO-2底物是DMA拓扑异构酶II（topoll），众所周知，该蛋白酶体在组织有丝分裂染色体方面具有重要功能。 Sumoylation的抑制会改变Topoll的染色体关联状态。我假设易生介导的有丝分裂液具有通过其底物调节有丝分裂染色体的组织的重要功能，该底物在后期的染色体分离中起着至关重要的作用。我建议通过确定Topoll Sumoylation（AIM2）的功能，并通过将新型的Piasy底物（AIM3）确定有丝分裂Sumoylation的调节机制，以及有丝分裂sumoylation的结果，并将其使用Xenopus egg提取物作为模型系统。该研究将证明SUMO-2修饰在脊椎动物中有丝分裂染色体的组织中的功能。确定邻近介导的Sumoylation的调节机制将提供一份宝贵的信息，以了解对各种细胞生理功能产生影响的Sumoylation途径。