Molecular Determinants of Chromosome Transmission and Cell Cycle Regulation

染色体传递和细胞周期调节的分子决定因素

基本信息

批准号：
8349186
负责人：
Munira Basrai
金额：
$ 129.04万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/8349186
关键词：
Affect Aging Aneuploidy Antineoplastic Agents Apoptosis Area Biochemical Biological Models Cell Cycle Cell Cycle Arrest Cell Cycle Checkpoint Cell Cycle Regulation Cell Death Cellular biology Centromere Chromatin Chromosome Segregation Chromosomes Collection Colorectal Cancer Complement Congenital Abnormality Copper DNA DNA Damage DNA Replication Damage Defect Deposition Disease Drosophila genus Drug Delivery Systems Ensure Eukaryota Failure Familial Amyotrophic Lateral Sclerosis Fission Yeast Gene Deletion Genes Genetic Genetic Materials Genetic Screening Genome Genome Stability Genomic Instability Genomics Goals Histone H3 Histone H4 Histones Homologous Gene Human Kinetochores Laboratories Lead Life Light Link Maintenance Malignant Neoplasms Mammalian Cell Mediating Mitotic Mitotic Checkpoint Molecular Molecular Chaperones Monitor Mutation Nature Organism Orthologous Gene Oxidative Stress Pathway interactions Phenotype Physiological Play Post-Translational Protein Processing Process Proteins Reactive Oxygen Species Recovery Research Research Project Grants Robot Role SOD1 gene Saccharomyces cerevisiae Saccharomycetales System Time Tumor Suppressor Genes Universities Variant Yeasts centromere protein A chromatin modification dosage gain of function human disease mutant novel overexpression prevent repaired response stoichiometry transmission process

项目摘要

We have used chromosome transmission fidelity (ctf) mutants and the deletion strain collections of S. cerevisiae to identify and characterize genes required for kinetochore function and checkpoint function. Studies with the ctf mutants led to the identification and characterization of the roles of SPT4 and NUP170 in chromosome segregation and spindle assembly checkpoint (SAC) function. We established a novel role for Spt4p in heterochromatic silencing. Using a cross-species approach, we showed that the yeast spt4 strains are complemented by human SPT4. Most importantly, we showed that S. cerevisiae SPT4 contributes to the proper localization of evolutionarily conserved centromeric histone H3 variant (CenH3) Cse4p. The major research goal of our laboratory is to investigate the molecular mechanisms that regulate the function of Cse4p and its interacting partners (Scm3p and Histone H4) to mediate faithful chromosome segregation. We investigated the mechanism of Cse4p localization and have established that mislocalization of Cse4p and altered histone stoichiometry lead to defects in chromosome transmission. Our studies have also shown that overexpression Scm3p and its human homolog HJURP leads to genome instability in yeast and human systems. We examined the effect of chromatin modifiers and post-translational modification of kinetochore proteins on the assembly/function of CenH3 chromatin. Our results showed that hypoacetylation state of centromeric histone H4 is critical for faithful chromosome segregation. Our recent results with Cse4p localization and histone dosage in S. cerevisiae are consistent with those in S. pombe, suggesting conservation of the underlying mechanisms. Thus, studies in S. cerevisiae that elucidate a mechanism for Cse4p localization and the role of chromatin modifications in centromere function may help us understand analogous pathways in humans and other systems. Our research on the molecular determinants of faithful chromosome transmission in S. cerevisiae will help us understand analogous processes in humans and their implications in human disease. Our laboratory is uniquely poised to utilize conventional genetic, biochemical, and cell biology approaches, as well as high-throughput genomic analysis for our research projects. We use an array of gene-deletion strains and a colony-picking robot for the identification of possible cancer drug targets and also for genetic screens by synthetic genome analysis (SGA), developed in the laboratory of Charlie Boone (University of Toronto).

我们已经使用染色体传递保真度（CTF）突变体和酿酒酵母的缺失应变收集来识别和表征Kinetochore功能和检查点功能所需的基因。使用CTF突变体的研究导致SPT4和NUP170在染色体分离和纺锤体组装检查点（SAC）功能中的作用鉴定和表征。我们确定了SPT4P在异质沉默中的新作用。使用跨物种方法，我们表明酵母SPT4菌株与人类SPT4相辅相成。最重要的是，我们表明酿酒酵母SPT4有助于适当地定位进化保守的中心透粒组蛋白H3变体（CENH3）CSE4P。我们实验室的主要研究目的是研究调节CSE4P及其相互作用伙伴（SCM3P和组蛋白H4）功能的分子机制，以介导忠实的染色体隔离。我们研究了CSE4P定位的机理，并确定CSE4P的错误定位和组蛋白化学计量的变化导致染色体传递中的缺陷。我们的研究还表明，过表达的SCM3P及其人类同源性Hjurp导致酵母和人类系统中的基因组不稳定性。我们检查了染色质修饰剂和动力学蛋白对CENH3染色质组装/功能的翻译后修饰的影响。我们的结果表明，丝粒组蛋白H4的低乙酰化状态对于忠实的染色体分离至关重要。我们最近在酿酒酵母中的CSE4P定位和组蛋白剂量的结果与S. pombe中的剂量一致，这表明保护了基本机制。因此，在酿酒酵母中阐明了CSE4P定位机制的研究以及染色质修饰在丝粒功能中的作用可能有助于我们了解人类和其他系统中的类似途径。我们对酿酒酵母中忠实染色体传播的分子决定因素的研究将有助于我们了解人类的类似过程及其对人类疾病的影响。我们的实验室有独特的准备利用常规的遗传，生化和细胞生物学方法，以及我们的研究项目的高通量基因组分析。我们使用一系列基因缺失菌株和一个挑选菌落的机器人来鉴定可能的癌症药物靶标，并通过合成基因组分析（SGA）（多伦多大学）开发的合成基因组分析（SGA）来鉴定遗传筛查。