CHARACTERIZATION OF ESSENTIAL PROTEINS INVOLVED IN CHROMOSOME SEGREGATION

参与染色体分离的必需蛋白的表征

• 批准号: 7723753
• 负责人: Philip A. Hieter
• 金额: $ 0.67万
• 依托单位: UNIVERSITY OF WASHINGTON
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-09-01 至 2009-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7723753
• 关键词: Affinity Chromatography; Anaphase; Aneuploid Cells; Biological Process; Cell division; Chromosomal Instability; Chromosome Segregation; Chromosomes; Complex; Computer Retrieval of Information on Scientific Projects Database; DNA Damage; DNA Repair; Defect; Essential Genes; Exhibits; Funding; Genes; Genetic Screening; Genomics; Goals; Grant; Institution; Knowledge; Laboratories; Lead; Maintenance; Malignant Neoplasms; Mammals; Metaphase; Methods; Mitosis; Mutate; Normal Cell; Process; Protein Analysis; Proteins; Rate; Research; Research Personnel; Resources; Role; Saccharomyces cerevisiae; Saccharomycetales; Sister Chromatid; Source; System; Techniques; United States National Institutes of Health; Yeasts; chromosome loss; cohesin; cohesion; daughter cell; interest; mutant; novel; tumor progression

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Normal cell division requires that all the chromosomes are duplicated and separated equally into the two daughter cells. During mitosis, sister chromatids are held together by protein complexes known as cohesins (by a process known as sister chromatid cohesion), until the metaphase to anaphase transition. Defective cohesion can lead to aneuploid cells by an increased rate of gain or loss of chromosomes (known as chromosome instability (CIN)). In mammalian systems, CIN may contribute to cancer progression and is exhibited by a variety of cancers, yet the mechanism by which it occurs is poorly understood. The budding yeast Saccharomyces cerevisiae has been useful for identifying genes that are involved in maintenance of genomic integrity and conserved in mammals. A recent genetic screen of a Ts mutant resource in the Hieter laboratory yielded three novel essential genes that lead to CIN, cohesion defects and sensitivity to DNA damaging agents when mutated. In order to fully understand the biological function of the corresponding proteins, a primary goal of this project is to identify interacting proteins that may yield information about which specific biological process these proteins are involved in. The primary technique used will be tandem affinity purification (TAP) in order to isolate the proteins of interest as part of intact complexes and to identify interacting partners. The TAP method has been shown to be useful for isolating biologically active complexes of sufficient purity for mass spectrometric analysis and protein identification. The results obtained from TAP will not only expand our knowledge of protein interaction networks in yeast but will also provide a starting point to further characterize the role of these novel proteins in DNA repair and sister chromatid cohesion establishment and/or maintenance.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 正常细胞分裂要求将所有染色体复制并平均分成两个子细胞。在有丝分裂过程中，姐妹染色质剂被称为粘着蛋白的蛋白质复合物（通过称为姐妹染色单体内聚力的过程）将其持有，直到中期为后期过渡。 有缺陷的凝聚力会通过增加染色体的增益或损失（称为染色体不稳定性（CIN））而导致非整倍体细胞。 在哺乳动物的系统中，CIN可能有助于癌症的进展，并由各种癌症表现出来，但其发生的机制知之甚少。 酿酒酵母的萌芽酵母菌对鉴定涉及维持基因组完整性并在哺乳动物中保守的基因很有用。 Hieter实验室中TS突变资源的最新遗传筛选产生了三个新的基本基因，这些基因导致CIN，凝聚力缺陷和对DNA突变时对DNA损害剂的敏感性。为了充分了解相应蛋白质的生物学功能，该项目的主要目标是识别相互作用的蛋白质，这些蛋白可能会产生有关这些蛋白所涉及的特定生物学过程的信息。所使用的主要技术将是串联亲和力纯化（TAP），以隔离感兴趣的蛋白质作为Intact Complectes的一部分，并确定相互作用的相互作用伙伴。 TAP方法已被证明可用于分离具有足够纯度的生物活性复合物，用于质谱分析和蛋白质鉴定。 从TAP获得的结果不仅将扩大我们对酵母中蛋白质相互作用网络的了解，而且还将提供一个起点，以进一步表征这些新型蛋白质在DNA修复中的作用和姐妹染色体凝聚建立和/或维持。