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APLF促进了跨链DNA交联修复和复制叉保护,以赋予西斯普拉丁耐药性

Cheng-Kuei Wu¹, Jia-Lin Shiu², Chao-Liang Wu³

¹Institute of Clinical Medicine, College of Medicine, National Cheng Kung University, No. 35, Xiao-Tong Road, Tainan 704, Taiwan.

Nucleic acids research

|March 23, 2024

概括

阿普拉和多核酸激酶/酸酶样因子 (APLF) 保护停滞的DNA复制分叉,并有助于跨链DNA交联修复. APLF 枯竭使癌细胞对西斯素敏感,突出显示了它在化学抵抗中的作用.

科学领域:

分子生物学分子生物学
癌症研究癌症研究
DNA 修复机制的修复机制

背景情况:

复制压力可能会阻碍DNA分叉,而分叉反转保护了这些分叉,以防止退化和DNA双链断裂 (DSB).
这种分叉保护机制矛盾的是,有助于癌细胞的化学抵抗.
聚 ((ADP-ribose) 聚合酶1 (PARP1) 被停滞的分叉激活并促进分叉逆转,而阿普拉和多核酸激酶/酸酶样因子 (APLF) 结合PARP1并以其在非同源端连接 (NHEJ) 中的作用而闻名.

研究的目的:

确定APLF在DNA跨链 (ICL) 修复和DNA分叉保护中的新功能.
阐明APLF在招募FANCD2到停滞的复制分叉中的作用.
调查APLF对癌细胞中西斯普拉丁耐药性的贡献.

主要方法:

细胞测试以评估DNA分叉稳定性和修复.
使用遗传方法 (例如,siRNA或CRISPR) 耗尽APLF.
分析蛋白质招募 (例如,APLF,PARP1,FANCD2) 停滞的叉使用技术,如免疫光或西方涂抹.

主要成果:

PARP1活动对于招募APLF到停滞不前的复制叉来说至关重要.
APLF促进了FANCD2的随后招募到停滞的分叉.
耗尽APLF损害了ICL修复,减少了FANCD2的招募,并导致MRE11核酶的新生DNA降解.
抗西斯普拉丁的癌细胞表现出升高的APLF水平和同源重组相关基因的增加表达.
耗尽APLF使细胞对思素敏感,并导致叉子的不稳定性.

结论:

APLF在促进跨链DNA交联修复和保护停滞不前的DNA复制分叉方面发挥了新而关键的作用.
APLF有助于癌细胞中对西斯普拉丁耐药的表型的发展.
准APLF可能是克服癌症治疗中西斯普拉丁耐药性的治疗策略.

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