...
首页> 外文期刊>Journal of Molecular Biology >An Mcm10 Mutant Defective in ssDNA Binding Shows Defects in DNA Replication Initiation
【24h】

An Mcm10 Mutant Defective in ssDNA Binding Shows Defects in DNA Replication Initiation

机译:Mcm10突变体的ssDNA结合缺陷显示DNA复制起始中的缺陷。

获取原文
获取原文并翻译 | 示例
   

获取外文期刊封面封底 >>

       

摘要

Mcm10 is an essential protein that functions to initiate DNA replication after the formation of the replication fork helicase. In this manuscript, we identified a budding yeast Mcm10 mutant (Mcm10-m2,3,4) that is defective in DNA binding in vitro. Moreover, this Mcm10-m2,3,4 mutant does not stimulate the phosphorylation of Mcm2 by Dbf4-dependent kinase (DDK) in vitro. When we expressed wild-type levels of mcm10-m2,3,4 in budding yeast cells, we observed a severe growth defect and a substantially decreased DNA replication. We also observed a substantially reduced replication protein A- chromatin immunoprecipitation signal at origins of replication, reduced levels of DDK-phosphorylated Mcm2, and diminished Go, Ichi, Ni, and San (GINS) association with Mcm2-7 in vivo. mcm5-bobl bypasses the growth defect conferred by DDK-phosphodead Mcm2 in budding yeast. However, the growth defect observed by expressing mcm10-m2,3,4 is not bypassed by the mcm5-bobl mutation. Furthermore, origin melting and GINS association with Mcm2-7 are substantially decreased for cells expressing mcm10-m2,3,4 in the mcm5-bobl background. Thus, the origin melting and GINS Mcm2-7 interaction defects we observed for mcm10-m2,3,4 are not explained by decreased Mcm2 phosphorylation by DDK, since the defects persist in an mcm5-bobl background. These data suggest that DNA binding by Mcm10 is essential for the initiation of DNA replication. (C) 2016 Elsevier Ltd. All rights reserved.
机译:Mcm10是必需的蛋白质,在复制叉解旋酶形成后,具有启动DNA复制的功能。在此手稿中,我们鉴定了出芽的酵母Mcm10突变体(Mcm10-m2,3,4),该突变体在体外DNA结合中存在缺陷。此外,该Mcm10-m2,3,4突变体在体外不刺激Dbf4依赖性激酶(DDK)引起的Mcm2磷酸化。当我们在萌芽的酵母细胞中表达mcm10-m2,3,4的野生型水平时,我们观察到严重的生长缺陷和DNA复制的大幅减少。我们还观察到复制起点的复制蛋白A染色质免疫沉淀信号大大降低,DDK磷酸化的Mcm2水平降低,并且体内的Go,Ichi,Ni和San(GINS)与Mcm2-7的关联减少。 mcm5-bob1绕开了DDK-磷脂酶Mcm2在发芽酵母中产生的生长缺陷。但是,通过表达mcm10-m2,3,4观察到的生长缺陷并没有被mcm5-bobl突变所绕过。此外,对于在mcm5-bobl背景中表达mcm10-m2、3、4的细胞,起源融化和与Mcm2-7的GIS关联显着降低。因此,我们观察到的针对mcm10-m2,3,4的原点熔解和GINS Mcm2-7相互作用缺陷不能通过DDK降低的Mcm2磷酸化来解释,因为缺陷持续存在于mcm5-bobl背景中。这些数据表明,Mcm10与DNA的结合对于DNA复制的启动至关重要。 (C)2016 Elsevier Ltd.保留所有权利。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号