Mechanism of Mitotic Checkpoint in Mammalian Cells

我的实验室的长期目标是获得对非整倍体在人类基因组中的作用的机械性见解 肿瘤发生。我们目前的重点是研究关键的机械和监管事件，这些事件说明 人类细胞中准确的染色体分离对于理解 致癌机制和杀灭癌细胞的新方法的开发。我们 专注于鉴定和表征动粒的分子组成，作为主要[ 努力了解准确的染色体分离的分子要求。这个 这项提议的目标是研究这些蛋白质中的一些是如何共同作用的，以形成具有功能的 连接着动粒的动粒：微管与有丝分裂检查点途径的相互作用。我们会 使用分子、生化和微观方法来实现我们的目标。这项提议将 重点研究进化保守的检查点蛋白激酶，hBUB 1和MPS 1，以及cdc27亚单位ot 后期促进复合体(APC)。我们将研究hBUB 1指定 动粒亚域的组装，由检查点蛋白hBUBR1，MAD1， Mad2和Cdc20。这些研究将是理解动态性质的关键 检查点蛋白和动点蛋白之间的相互作用。我们将描述HMPS 1的作用 通过检测有丝分裂检查点通路中有丝分裂检查点通路在着丝点和细胞周期中的重要性 将信号从未连接的动控中心传送到APC。最后，我们制作了这部小说 发现APC的CDC27亚单位促进了对两者APC的检查点抑制 人类和萌芽中的酵母。我们对CDC27如何做到这一点的分析提供了一个独特的“底部-- UP“方法来研究信号通路，它允许细胞即使只有一个独立的 来自过早退出有丝分裂的染色体。 性能 关键人员。 从本金开始 名字 站点(S)(组织、市、州) 宾夕法尼亚州费城福克斯·蔡斯癌症中心癌症研究所 请参阅说明。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，姓氏在前。 组织 格式如下所示。 在项目中的角色 首页--期刊主要分类--期刊细介绍 首页--期刊主要分类--期刊细介绍 福克斯大通癌症中心月春癌症研究所 马库斯，福克斯·蔡斯癌症中心亚当癌症研究所 梅洛伊，帕特里夏癌症研究所，福克斯·蔡斯癌症中心 首席调查员 联合调查员 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 小灵通398(05/01版)第_页 在整个应用程序的底部连续编号页码。不要使用后缀，如3a、3b、 表单第2页 2. 首席调查员/项目主任(最后、第一、中间)：严，L 打印页面顶部和后续页面的theprincipalinvestigatorprogradmirectormustbeprovidedat名称。 研究补助金 目录 页码_ 1 主页....................................................................................................................................................................................... 2. 描述，

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,