人类RAD18对细胞类型特定的超重组抑制与PCNA K164无处不在化有关
bioRxiv : the preprint server for biology
|September 16, 2024
概括
在RAD18中使用RAD18.
科学领域:
- DNA 修复机制的修复机制
- 细胞对DNA损伤的反应
背景情况:
- 同源重组 (HR) 和转化合成 (TLS) 对于DNA复制和修复至关重要.
- RAD18是一种E3泛素酶,涉及HR和TLS通路.
- 已知RAD18在通过PCNA无处不在的TLS中的作用,但其在HR中的作用不太清楚.
研究的目的:
- 研究RAD18在人类细胞系的同类重组 (HR) 和转化合成 (TLS) 中的作用.
- 要确定RAD18在这些通路中的功能是否取决于细胞类型.
- 为了阐明RAD18,PCNA无化和重组之间的联系.
主要方法:
- 在HCT116,hTERT RPE-1和DLD1人类细胞系中RAD18的失活.
- 对姐妹染色体交换,基因转换和基因向进行分析,以评估重组.
- 转化合成 (TLS) 活性和克隆原生存活率的评估.
- 产生PCNA K164突变体和UBC9敲击,以研究无处化和SUMOylation.
主要成果:
- 在HCT116细胞中,RAD18的丧失导致了超重组因性,但在hTERT RPE-1或DLD1细胞中没有.
- 在没有降低存活率的情况下,RAD18缺乏会损害HCT116细胞中的TLS活性.
- PCNA K164无处不在突变体表现出类似的超重组生成和减少的TLS表型.
- 阻止PCNA K164 SUMOylation并没有影响过度重组.
结论:
- 人类RAD18在不同的DNA填补途径中的作用是特定于细胞类型的.
- 这些功能与RAD18介导的PCNA K164无化密切相关,而不是SUMOylation.
- RAD18影响了复合和TLS,结果因细胞系而异.
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