在S.S.中识别1600个复制源. 类植物
Eric J Foss1, Carmina Lichauco1, Tonibelle Gatbonton-Schwager1
1Clinical Research Division, Fred Hutch Cancer Center, Seattle, WA 98109.
bioRxiv : the preprint server for biology
|November 28, 2023
概括
研究人员发现,酵母中的DNA复制起源至少比以前认为的多三倍. 这一发现表明,酵母DNA复制与人类复制更相似,而不是以前所认为的.
科学领域:
- 分子生物学分子生物学
- 遗传学 是一个遗传学.
- 细胞生物学 细胞生物学
背景情况:
- 酵母基因组有大约500个已知的DNA复制起源,启动机制是众所周知的.
- 复制能力是通过在G1阶段加载Mcm复制酶来实现的.
- 之前的研究通过Mcm结合定义了起源,通常是ARS共识序列 (ACS) 附近的一个单双六合体.
研究的目的:
- 为了识别整个酵母基因组中的所有Mcm结合点.
- 为了确定Mcm结合点在DNA复制启动中的功能意义.
- 重新评估酵母中DNA复制起源的总数和分布.
主要方法:
- 利用染色体内源性裂变 (CEC) 通过将Mcm子单元融合到微球菌核酶.
- 分析了全基因组的Mcm结合点信号强度 (超过3个数量级).
- 在S阶段与单链DNA (ssDNA) 生产相关的Mcm丰度和分析的原始标志 (无核素体区域,ACS,GC偏差).
主要成果:
- 确定了成千上万的Mcm结合点,在前1600个中具有显著的复制活性.
- 5,500个最丰富的Mcm网站显示复制起源的标志,包括特定的基因组定位和侧面序列.
- 与ACS相比,在Mcm双六合体负载中表现出方向偏差,与体外Orc活动一致.
结论:
- 酵母中的DNA复制起源至少比以前估计的多3倍.
- 复制可能在几乎所有基因间区域开始,挑战离散的,有限的起源的概念.
- 酵母复制起源的更广泛分布表明,与人类复制过程的相似性比以前假设的更大.
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