Comprehensive Mutational Analysis of the BRCA1-Associated DNA Helicase and Tumor-Suppressor FANCJ/BACH1/BRIP1.
Comprehensive Mutational Analysis of the BRCA1-Associated DNA Helicase and Tumor-Suppressor FANCJ/BACH1/BRIP1.
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DOI：
10.1158/1541-7786.mcr-20-0828
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发表时间：
2021-06
期刊：
影响因子：
--
通讯作者：
Cantor SB
中科院分区：
文献类型：
--
作者：
Calvo JA;Fritchman B;Hernandez D;Persky NS;Johannessen CM;Piccioni F;Kelch BA;Cantor SB
FANCJ(BRIP1/BACH1)是一种遗传性乳腺癌和卵巢癌(HBOC)基因，编码DNA解旋酶。与HBOC基因BRCA1和BRCA2相似，FANCJ在处理顺铂等化疗药物诱导的DNA链间交联(ICL)过程中起着关键作用。因此，缺乏FANCJ或其催化活性的细胞对ICL诱导剂敏感。不幸的是，大多数FANCJ临床突变仍然没有特征性，限制了有效使用顺铂治疗突变FANCJ肿瘤的治疗机会。在这里，我们试图进行一次全面的筛查，以确定FANCJ功能丧失(LOF)突变。我们建立了一个代表约450个患者来源的FANCJ无义和错义突变的FANCJ慢病毒突变文库，以将FANCJ突变引入FANCJ基因敲除(K/O)HeLa细胞。我们进行了高通量筛选来鉴定FANCJ LOF突变体，与野生型FANCJ相比，这些突变体未能有力地恢复对ICL诱导剂顺铂或丝裂霉素C(MMC)的抗性。基于对顺铂或丝裂霉素耐药的失败，我们鉴定了26个错义和25个无义LOF突变。无义突变阐明了截断位置和ICL敏感性之间的关系，因为在氨基酸860之前的大多数无义突变赋予ICL敏感性。LOF突变子集的进一步验证证实了该筛查能够识别无法赋予ICL耐药性的FANCJ突变。最后，将LOF突变的位置映射到新的同源模型中，以获得额外的功能信息。
FANCJ (BRIP1/BACH1) is a hereditary breast and ovarian cancer (HBOC) gene encoding a DNA helicase. Similar to HBOC genes, BRCA1 and BRCA2, FANCJ is critical for processing DNA inter-strand crosslinks (ICL) induced by chemotherapeutics, such as cisplatin. Consequently, cells deficient in FANCJ or its catalytic activity are sensitive to ICL-inducing agents. Unfortunately, the majority of FANCJ clinical mutations remain uncharacterized, limiting therapeutic opportunities to effectively use cisplatin to treat tumors with mutated FANCJ. Here we sought to perform a comprehensive screen to identify FANCJ loss-of-function (LOF) mutations. We developed a FANCJ lentivirus mutation library representing ~450 patient derived-FANCJ nonsense and missense mutations to introduce FANCJ mutants into FANCJ knockout (K/O) HeLa cells. We performed a high-throughput screen to identify FANCJ LOF mutants that, as compared to wildtype FANCJ, fail to robustly restore resistance to ICL-inducing agents, cisplatin or mitomycin C (MMC). Based on the failure to confer resistance to either cisplatin or MMC, we identified 26 missense and 25 nonsense LOF mutations. Nonsense mutations elucidated a relationship between location of truncation and ICL sensitivity, as the majority of nonsense mutations before amino acid 860 confer ICL sensitivity. Further validation of a subset of LOF mutations confirmed the ability of the screen to identify FANCJ mutations unable to confer ICL resistance. Finally, mapping the location of LOF mutations to a new homology model additional functional information.
布卢姆综合征蛋白的精氨酸指：其结构组织及其在能量耦合中的作用
影响因子：
14.9
作者：
Ren H;Dou SX;Rigolet P;Yang Y;Wang PY;Amor-Gueret M;Xi XG
通讯作者：
Xi XG
通过增加的运动ATPase或负载链的长度来克服骨架不连续性对Bach1（FANCJ）解旋酶的抑制。
影响因子：
14.9
作者：
Gupta, Rigu;Sharma, Sudha;Doherty, Kevin M;Sommers, Joshua A;Cantor, Sharon B;Brosh, Robert M Jr
通讯作者：
Brosh, Robert M Jr