调节的真核DNA复制起源与纯化蛋白质一起燃烧
Joseph T P Yeeles1, Tom D Deegan1, Agnieszka Janska1
1Cancer Research UK London Research Institute, Clare Hall Laboratories, South Mimms EN6 3LD, UK.
Nature
|March 6, 2015
概括
研究人员使用42种蛋白质重建了真核细胞DNA复制启动. 这个过程受到循环素依赖激酶 (CDK) 和Dbf4依赖激酶 (DDK) 的严格调节,以确保精确的基因组重复.
科学领域:
- 分子生物学分子生物学
- 细胞生物学 细胞生物学
- 生物化学 生物化学
背景情况:
- 细胞DNA复制从多个起源开始,需要对基因组复制进行精确的调节.
- 复制启动发生在两个阶段:小染色体维护 (MCM) 复杂加载在G1中,然后在S阶段激活到CMG酶.
研究的目的:
- 复制和分析在体外调节DNA复制启动的最小蛋白质要求.
- 阐明循环素依赖激酶 (CDK) 和Dbf4依赖激酶 (DDK) 在控制复制启动中的作用.
主要方法:
- 使用16个纯化的复制因子 (42个多) 重建芽酵母DNA复制启动.
- 在体外测试以观察来源依赖的启动和调节机制.
主要成果:
- 重建后的系统在体内总结了DNA复制启动的调节.
- 通过化ORC,CDK抑制了MCM的加载,通过化Sld2和Sld3促进了CMG的形成.
- DDK通过酸化MCM促进复制,与CDK相比,时间灵活.
结论:
- 定义了最小的蛋白质补充物,酶基质,以及对于调节的真核生物DNA复制必不可少的辅因子.
- 证明了CDK和DDK在控制复制启动时间和进展方面的独特监管作用.
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