CHROMOSOME MOVEMENT IN PROMETAPHASE

前期染色体运动

• 批准号: 2188245
• 负责人: GARY J. GORBSKY
• 金额: $ 11.69万
• 依托单位: UNIVERSITY OF VIRGINIA
• 依托单位国家: 美国
• 财政年份: 1994
• 资助国家: 美国
• 起止时间: 1994-01-01 至 1996-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2188245
• 关键词: antibody specificity; biological signal transduction; cell cycle; centromere; chromosome movement; gene expression; genetic manipulation; immunofluorescence technique; microinjections; mitotic spindle apparatus; phosphoprotein phosphatase; phosphorylation; polymerase chain reaction; protein kinase; protein purification; protein structure; western blottings

DESCRIPTION (adapted from applicant's abstract): The kinetochore mediates the movement of chromosomes along the microtubules of the mitotic spindle. The Cyert antibody, which is directed to an incompletely characterized, phosphorylated epitope, binds to the kinetochores of chromosomes in prometaphase in a unprecedented and intriguing manner. Chromosomes moving toward the metaphase plate express the phospho-epitope strongly on the leading kinetochore but weakly on the trailing kinetochore. This differential expression of the epitope may reflect a biochemical pathway regulating chromosome movement and the progression of mitosis. Two competing hypotheses for the molecular nature of the epitope recognized by the Cyert antibody will be tested. First, the antibody recognizes a regulatory phosphorylation on a protein that powers or guides chromosome movement. Second, the epitope recognizes an enzymatic intermediate of a phosphatase that is differentially regulated at kinetochores. The aims of this application are: (1) to examine how expression of the phospho-epitope changes in relation to chromosome movement and mitotic progression; (2) to determine how kinases and phosphatases regulate that expression; (3) to identify the proteins that contain the epitope; and (4) to understand how positional and mechanical signals in the mitotic spindle are converted to biochemical information that regulates chromosome movement and progression through mitosis. These aims will be achieved by immunochemical isolation, fractionation, and molecular characterization of kinetochore proteins. These efforts will be complemented by the analysis of chromosome behavior in live cells, immunolabeling studies, micromanipulation of chromosomes, and the microinjection of antibodies and inhibitors.

描述（改编自申请人的摘要）：动粒 介导染色体沿着微管的运动 有丝分裂纺锤体。 Cyert 抗体，针对 不完全表征的磷酸化表位，与 染色体着丝粒在前中期以前所未有的方式 有趣的方式。 染色体向中期板移动 在主导着丝粒上强烈表达磷酸表位，但 尾随动粒上微弱。 这种差异表达 表位可能反映调节染色体运动的生化途径 和有丝分裂的进展。 两个相互竞争的假设 Cyert 抗体识别的表位的分子性质将 被测试。 首先，抗体识别调节性磷酸化 驱动或引导染色体运动的蛋白质。 其次， 表位识别磷酸酶的酶中间体，即 着丝粒受到差异性调节。 此应用程序的目的 是：（1）检查磷酸表位的表达如何变化 与染色体运动和有丝分裂进展的关系； (2) 至 确定激酶和磷酸酶如何调节该表达； (3) 至 识别含有表位的蛋白质； (4) 理解 有丝分裂纺锤体中的位置和机械信号如何 转化为调节染色体运动的生化信息 并通过有丝分裂进展。 这些目标将通过以下方式实现 免疫化学分离、分级分离和分子表征 动粒蛋白。 这些努力将得到补充 活细胞染色体行为分析、免疫标记研究、 染色体显微操作和抗体显微注射 和抑制剂。