Identification of Molecular Pathways in Cancer Biology

癌症生物学中分子途径的鉴定

• 批准号: 6817125
• 负责人: MICHAEL B YAFFE
• 金额: $ 14.4万
• 依托单位: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2006-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6817125
• 关键词: DNA damage; Escherichia coli; SDS polyacrylamide gel electrophoresis; biological signal transduction; cell growth regulation; cell proliferation; complementary DNA; cyclin dependent kinase; enzyme activity; expression cloning; genetic library; high throughput technology; method development; microorganism culture; neoplastic cell; phosphopeptides; phosphorylation; protein binding; protein structure function; reagent /indicator; serine threonine protein kinase

DESCRIPTION (provided by applicant): Controlling the onset and progression of mitotic cell division is critical to prevent human cancer. Much of this regulation involves the timely activation and inactivation of cyclin-dependent protein kinases that regulate progression through the cell cycle. In addition, cells must ensure that their genetic information is accurately transmitted to each daughter cell by activating a series of DNA damage-response kinases to halt the cell cycle in response to genotoxic stress. Many details of the molecular events that occur downstream of mitotic and DNA damage-response kinases, however, are unknown. In this regard, phosphoSerine/Threonine (pSer/pThr)-binding domains such as 14-3-3 proteins, FHA domains, WW domains, and WD40 repeats are increasingly emerging as critical doma ns that link upstream kinase signaling cascades to downstream effector functions by directly mediating the phosphorylation-dependent formation of multi-protein signaling complexes. The long-term goals of the applicant are to identify and characterize (pSer/pThr)-binding domains involved in regulating cell proliferation. In this proposal we seek funds to develop and improve a new phosphopeptide library-based proteomic screening technology that simultaneously: (1) reveals specific pSer/pThr-binding domains downstream of cyclin-dependent kinases and DNA damage-responsive kinases; (2) allows determination of the optimal sequence motifs recognized by the newly-identified domain; (3) provides reagents for biophysical, cell biological, and structural studies of the function of the newly identified domain in regulating mitotic progression, as well as reagents for high-throughput screening for discovery of small molecule inhibitors; and (4) facilitates bioinformatics and systems biology efforts to identify specific kinase targets within signaling pathways that mediate cell cycle control. This technology will have a major impact on proteomicsand bioinformatics/systems-biology based approaches to define kinase-dependent molecular pathways in cancer biology and will identify new therapeutic targets for drug design.

描述（由申请人提供）：控制有丝分裂细胞分裂的发生和进展对于预防人类癌症至关重要。这种调节大部分涉及细胞周期蛋白依赖性蛋白激酶的及时激活和失活，这些蛋白激酶调节细胞周期的进展。此外，细胞必须通过激活一系列 DNA 损伤反应激酶来停止细胞周期以响应基因毒性应激，从而确保其遗传信息准确地传递到每个子细胞。然而，有丝分裂和 DNA 损伤反应激酶下游发生的分子事件的许多细节尚不清楚。在这方面，磷酸丝氨酸/苏氨酸 (pSer/pThr) 结合域，例如 14-3-3 蛋白、FHA 域、WW 域和 WD40 重复序列，日益成为连接上游激酶信号级联与下游效应子功能的关键域。通过直接介导多蛋白信号复合物的磷酸化依赖性形成。申请人的长期目标是鉴定和表征参与调节细胞增殖的(pSer/pThr)结合域。在本提案中，我们寻求资金来开发和改进一种新的基于磷酸肽库的蛋白质组筛选技术，该技术同时：（1）揭示细胞周期蛋白依赖性激酶和 DNA 损伤响应激酶下游的特定 pSer/pThr 结合域； (2) 允许确定新识别的结构域识别的最佳序列基序； (3) 提供用于新鉴定的结构域在调节有丝分裂进展中的功能的生物物理、细胞生物学和结构研究的试剂，以及用于发现小分子抑制剂的高通量筛选的试剂； (4) 促进生物信息学和系统生物学工作，以确定介导细胞周期控制的信号通路中的特定激酶靶点。该技术将对基于蛋白质组学和生物信息学/系统生物学的方法产生重大影响，以定义癌症生物学中激酶依赖性分子途径，并将确定药物设计的新治疗靶点。