利用酵母双杂交系统筛选并鉴定出与MCM4 C末端结构域（MCM4-CTD）相互作用的蛋白

发布时间：2018-03-14 14:55

本文选题：微小染色体维持蛋白复合物　切入点：MCM4-CTD　出处：《华中科技大学》2012年硕士论文　论文类型：学位论文

【摘要】：DNA复制的严谨性和准确性对于遗传信息的传递有重要的作用，，微小染色体维持蛋白复合物（MCM complex）是由六个亚基组成的六聚体，在真核细胞的DNA复制过程中起解旋酶的作用。在DNA损伤（DNA damage）时，复制叉会停止，过度的解旋会产生过多的单链DNA（ssDNA）从而能引起复制叉及整个基因组的不稳定。研究发现，MCM复合物中的一个最保守的亚基MCM4的C末端结构域（MCM4-CTD），其编码基因的大小为648bp，可以调控整个MCM复合物，在停止的复制叉处阻止它过度解旋，防止产生过多的单链DNA，这就预示着在DNA损伤时存在DNA损伤反应过程中相关的MCM4-CTD的重要作用。我们利用酵母双杂交实验，将MCM4-CTD克隆到BD载体上，从酵母的cDNA文库中筛选并鉴定出与之相互作用的蛋白SNF12，它是染色体重塑蛋白复合物（chromatin remodeling complex）SWI/SNF中的一个亚基。由此我们推测出在DNA损伤反应的过程中，染色体的重塑活性也参与到DNA复制的调控中。
[Abstract]:The rigour and accuracy of DNA replication play an important role in the transmission of genetic information. The microchromosome maintenance protein complex (MCM complex) is a hexamer composed of six subunits. It acts as a helicase in the process of DNA replication in eukaryotic cells. During DNA damage, the replication fork stops. Excessive unwinding produces excessive single-stranded DNA ssDNA), which can lead to replication forks and instability of the entire genome. It has been found that the C-terminal domain of MCM4, one of the most conserved subunits of the complex, is MCM4-CTDN, which encodes a gene size of 648 bp. Can regulate the entire MCM complex, We use yeast two-hybrid experiments to prevent excessive unwinding and excessive production of single strand DNAs at the stop replication forks, which indicate the important role of MCM4-CTD in the process of DNA damage response during DNA damage. MCM4-CTD was cloned into BD vector, and the interacting protein SNF12 was screened and identified from yeast cDNA library, which is a subunit of chromosome remodeling protein complex remodeling complex)SWI/SNF. We speculate that SNF12 is involved in the process of DNA damage response. The activity of chromosome remodeling is also involved in the regulation of DNA replication.
【学位授予单位】：华中科技大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R3416

【共引文献】