INTERACTIONS OF THE MCM PROTEINS AT REPLICATION ORIGINS

MCM 蛋白在复制起点的相互作用

基本信息

批准号：
6519162
负责人：
BIK-KWOON TYE
金额：
$ 34.58万
依托单位：
CORNELL UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1978
资助国家：
美国
起止时间：
1978-04-01 至 2004-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6519162
关键词：
DNA footprinting DNA replication origin Saccharomyces cerevisiae cell cycle cell cycle proteins fungal genetics fungal proteins gene mutation protein protein interaction transcription factor

项目摘要

The initiation of DNA synthesis is a complex multi-step process that requires the coordinated actions of a number of replication initiation factors. This process begins with the ordered assembly of a pre- replication complex (pre-RC) at replication origins during the G1 phase. Studies in Saccharomyces cerevisiae indicate that the pre-RC consists of evolutionarily conserved replication initiation factors that include the origin recognition complex, ORC, a complex of sib subunits, and the Mcm2-7 proteins, a family of six homologous proteins. Initiation of DNA synthesis is activated by the successive phosphorylations of components of the pre-RC by at least two Cdks (cyclin dependent kinases) or Cdk- like kinases, Cdc28-Clb and Cdc-7-Dbf4. Initiation of DNA synthesis at replication origins is accompanied by a transition of the pre-RC to the elongation complex. As the elongation complex migrates away from the replication origin, the pre-RC is replaced by the post-replication complex (post-RC), which appears to include the ORC but not the other known components of the pre-RC. Previous studies showed that Mcm10 physically interacts with members of the Mcm2-7 proteins. We will investigate the multiple roles of Mcm10 in the assembly and disassembly of the pre-RC and its role in the migration of elongation forms in conjunction with the Mcm2-7 proteins. Significant efforts will also be devoted to the analysis of the functional relationship between Mcm10 and other components of the pre-RC. These studies should provide information about the mechanism that restricts DNA synthesis to once per cell cycle in eukaryotes. Cancer cells are characterized by their unregulated cell divisions. This study addresses a fundamental problem in the control of DNA replication and cell division using Saccharomyces cerevisiae as the model. Through understanding of normal cellular processes that regulate DNA replication, it may be possible to elucidate the molecular basis for abnormalities or defects that lead to the disease state in cancer cells.

DNA合成的起始是一个复杂的多步骤过程需要多个复制启动的协调动作因素。这个过程从预制件的有序组装开始 G1 期复制起始点的复制复合物（pre-RC）。对酿酒酵母的研究表明，前 RC 包括进化上保守的复制起始因子包括起源识别复合体，ORC，同胞亚基的复合体，以及 Mcm2-7 蛋白，一个由六种同源蛋白组成的家族。 DNA的起始合成是通过组分的连续磷酸化来激活的前 RC 至少由两个 Cdks（细胞周期蛋白依赖性激酶）或 Cdk- 如激酶、Cdc28-Clb 和 Cdc-7-Dbf4。 DNA合成起始于复制起点伴随着前 RC 到 RC 的转变伸长复合体。当伸长复合物从复制起点，前 RC 被复制后替换复合体（后 RC），其中似乎包括兽人但不包括其他人预 RC 的已知组件。先前的研究表明Mcm10 与 Mcm2-7 蛋白的成员发生物理相互作用。我们将研究Mcm10在组装和拆卸中的多种作用前RC的作用及其在伸长形式迁移中的作用与 Mcm2-7 蛋白结合。也将做出重大努力致力于分析Mcm10与预 RC 的其他组件。这些研究应该提供信息关于限制 DNA 合成每个细胞周期一次的机制在真核生物中。癌细胞的特点是细胞不受调控部门。这项研究解决了控制中的一个基本问题使用酿酒酵母作为 DNA 复制和细胞分裂模型。通过了解调节的正常细胞过程 DNA复制，或许可以阐明DNA复制的分子基础导致癌细胞疾病状态的异常或缺陷。