Mechanism of Mitotic Checkpoint in Mammalian Cells

我的实验室的长期目标是获得非整倍体的作用机制的见解， 肿瘤发生我们目前的重点是研究关键的机械和监管事件， 人类细胞中染色体的精确分离对于理解 研究导致癌症的机制和开发杀死癌细胞的新方法。我们 已经集中于鉴定和表征动粒的分子组分作为主要[ 努力了解准确的染色体分离的分子要求。的 这项提案的目标是研究这些蛋白质如何共同作用，使一个功能 连接动粒：微管相互作用与有丝分裂检查点通路的动粒。我们将 使用分子、生物化学和微观方法来实现我们的目标。这项建议会 重点是进化上保守的检查点激酶hBUB 1和MPS 1，以及Cdc 27亚基， 后期促进复合物（APC）我们将研究hBUB 1指定 由检查点蛋白hBUBR 1，MAD 1， MAD2和Cdc20。这些研究对于理解生物多样性的动态性质至关重要。 检查点蛋白和动粒之间的相互作用。我们将描述hMPS 1的作用 激酶在有丝分裂检查点途径中的重要性， 将信号从未附着的动粒转导至APC。最后，我们把小说 发现APC的Cdc27亚基促进APC在两种细胞中的检查点抑制， 人类和芽殖酵母我们对Cdc27如何实现这一点的分析提供了一个独特的"底部"- “向上”的方法来研究信号通路，即使是一个单独的细胞， 染色体过早退出有丝分裂。 性能 关键人员。 从Principal开始 名称 地点（组织、城市、州） 癌症研究所，Fox Chase癌症中心，费城，PA 参见说明。根据需要使用续页提供所需信息 调查员按字母顺序列出所有其他关键人员，姓在前。 组织 格式如下。 在项目中的作用 Yen，Timothy J.癌症研究所，福克斯蔡斯癌症中心 文森特·瓜奇福克斯蔡斯癌症中心癌症研究所 李玉春癌症研究所，福克斯蔡斯癌症中心 马库斯，亚当癌症研究所，福克斯蔡斯癌症中心 梅洛伊，帕特里夏癌症研究所，福克斯大通癌症中心 主要研究者 合作研究 博士后助理 博士后助理 博士后助理 披露许可声明仅适用于SBIPJSTTR。参见说明，r3是[]否 PHS 398（2001年5月修订版）第_页 在整个应用程序的底部连续编号页面。不要使用诸如3a，3b， 表格第2页 2 主要研究者/项目负责人（最后，第一，中间）：YEN，Timothy L 必须在打印页和每个续页的顶部提供首席执行官和首席执行官的姓名。 研究资助 目录 页码_ 1 首页....................................................................................................................................................................................... 2 说明，

The long-term objective of my lab is to obtain mechanistic insights into the role of aneuploidy in tumorogenesis. Our current focus on studying the key mechanical and regulatory events that specify accurate chromosome segregation in human cells is of direct importance towards understanding mechanisms that cause cancer and for the development of novel approaches to kill cancer cells. We have focused on identifying and characterizing the molecular components of the kinetochore as a majo[ effort towards understanding the molecular requirements for accurate chromosome segregation. The goals of this proposal are to examine how some of these proteins work together to make a functional kinetochore that links kinetochore:microtubule interactions to the mitotic checkpoint pathway. We will use molecular, biochemical and microscopic approaches to accomplish our goals. This proposal will focus on the evolutionary conserved checkpoint kinases, hBUB 1 and MPS 1, and the Cdc27 subunit ot the Anaphase Promoting Complex (APC). We will examine the mechanism by which hBUB 1 specifies the assembly of subdomain of the kinetochore that consists of checkpoint proteins hBUBR1, MAD1, MAD2 and Cdc20. These studies will be critical for understanding the dynamic nature of the interactions between checkpoint proteins and kinetochores. We will characterize the role of the hMPS 1 kinase in the mitotic checkpoint pathway by examining its importance at kinetochores and in transducing the signal from unattached kinetochores to the APC. Lastly, we have made the novel discovery that the Cdc27 subunit of the APC facilitates checkpoint inhibition of the APC in both humans and budding yeast. Our analysis of how Cdc27 accomplishes this provides a unique "bottoms- up" approach to study the signaling pathway that allows cells with even a single unattached chromosome from prematurely exiting mitosis. PERFORMANCE KEY PERSONNEL. Start with Principal Name SITE(S) (organization, city, state) Institute for Cancer Research, Fox Chase Cancer Center, Philadelphia, PA See instructions. Use continuation pages as neededto provide the required information Investigator. List all other key personnel in alphabetical order, last name first. Organization in the format shown below. Role on Project Yen, Timothy J. Institute for Cancer Research, Fox Chase Cancer Center Guacci, Vincent A. Institute for Cancer Research, Fox Chase Cancer Center Li, Yueh-chun Institute for Cancer Research, Fox Chase Cancer Center Marcus, Adam Institute for Cancer Research, Fox Chase Cancer Center Melloy, Patricia Institute for Cancer Research, Fox Chase Cancer Center Principal Investigator Co-Investigator Postdoctoral Associate Postdoctoral Associate Postdoctoral Associate Disclosure Permission Statement ApOlicable to SBIPJSTTR Only. See instructions, r3 Yes [] No PHS 398 (Rev. 05/01) Page ___ Number pages consecutively at the bottom throughout the application. Do not use suffixes such as 3a, 3b, Form Page 2 2 Principal Investigator/Program Director (Last, first,middle): YEN, Timothy L The nameof theprincipalinvestigatorprogradmirectormustbeprovidedat thetop ofeachprintedpageandeachcontinuationpage. RESEARCH GRANT TABLE OF CONTENTS Page Numbe_ 1 Face Page ....................................................................................................................................................................................... 2 Description,