Regulation of the kinetochore-microtuble interface

着丝粒-微管界面的调节

• 批准号: 6836576
• 负责人: WALTER E GALL
• 金额: $ 4.73万
• 依托单位: UNIV OF NORTH CAROLINA CHAPEL HILL
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-08-01 至 2007-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6836576
• 关键词: Xenopus; Xenopus oocyte; cell component structure /function; cell cycle; cell cycle proteins; cell free system; centromere; chromosome movement; confocal scanning microscopy; gene mutation; intermolecular interaction; microtubule associated protein; microtubules; molecular assembly /self assembly; phase contrast microscopy; phosphorylation; postdoctoral investigator; protein kinase; protein localization; protein structure function; site directed mutagenesis; time resolved data; tissue /cell culture

DESCRIPTION (provided by applicant): In mitosis, proper microtubule (MT) attachment to kinetochores is critical for faithful chromosome segregation. Our lab has shown that errors in kinetochore attachment occur at a high frequency in human tissue culture cells and these have to be corrected for accurate segregation. Budding yeast studies indicate a role for mitotic kinase Ipl1 (human Aurora B ortholog) in correcting MT misattachments at kinetochores. These studies show that Ipl1 phosphorylates the highly conserved outer kinetochore component Ndc80 (human Hec1 ortholog) and a MT-associated protein Dam1 (no known human ortholog). Genetic and mutagenesis studies indicate that Aurora B phosphorylation of these substrates weakens kinetochore interactions with kinetochore MTs (kMTs). Preliminary data from my yeast studies suggest Aurora B regulates another MT-associated protein Dim1 (human EB1). EB1 proteins are known to bind and promote stability of plus-end kMTs. I propose to investigate how human Hec1 and EB1 function are regulated by aurora B at the vertebrate kMT interface. I will examine the consequences of presence or absence of Aurora B phosphorylation of these k-MT substrates in mammalian cells.

描述（由申请人提供）： 在有丝分裂中，微管（MT）正确附着在着丝粒上对染色体的分离至关重要。我们的实验室已经表明，在人类组织培养细胞中，动粒附着的错误发生频率很高，必须对这些错误进行纠正以获得准确的分离。芽殖酵母的研究表明，有丝分裂激酶Ipl 1（人极光B直系同源物）的作用，在纠正MT错误附着在着丝粒。 这些研究表明，Ipl 1磷酸化高度保守的外动粒组件Ndc 80（人类Hec 1直系同源物）和MT相关蛋白Dam 1（没有已知的人类直系同源物）。遗传和诱变研究表明，极光B磷酸化这些底物减弱动粒与动粒MT（kMT）的相互作用。我的酵母研究的初步数据表明，极光B调节另一种MT相关蛋白Dim 1（人类EB 1）。已知EB 1蛋白结合正末端kMT并促进其稳定性。我建议研究人类Hec 1和EB 1的功能是如何调节极光B在脊椎动物的kMT接口。我将检查哺乳动物细胞中这些k-MT底物是否存在Aurora B磷酸化的后果。