National Institute on Aging, NIH, NIH Biomedical Research Center, Baltimore, MD, USA.
Mol Cell Biol. 2013 Jun;33(11):2212-27. doi: 10.1128/MCB.01256-12. Epub 2013 Mar 25.
FANCJ mutations are linked to Fanconi anemia (FA) and increase breast cancer risk. FANCJ encodes a DNA helicase implicated in homologous recombination (HR) repair of double-strand breaks (DSBs) and interstrand cross-links (ICLs), but its mechanism of action is not well understood. Here we show with live-cell imaging that FANCJ recruitment to laser-induced DSBs but not psoralen-induced ICLs is dependent on nuclease-active MRE11. FANCJ interacts directly with MRE11 and inhibits its exonuclease activity in a specific manner, suggesting that FANCJ regulates the MRE11 nuclease to facilitate DSB processing and appropriate end resection. Cells deficient in FANCJ and MRE11 show increased ionizing radiation (IR) resistance, reduced numbers of γH2AX and RAD51 foci, and elevated numbers of DNA-dependent protein kinase catalytic subunit foci, suggesting that HR is compromised and the nonhomologous end-joining (NHEJ) pathway is elicited to help cells cope with IR-induced strand breaks. Interplay between FANCJ and MRE11 ensures a normal response to IR-induced DSBs, whereas FANCJ involvement in ICL repair is regulated by MLH1 and the FA pathway. Our findings are discussed in light of the current model for HR repair.
FANCJ 突变与范可尼贫血症 (FA) 相关,并增加乳腺癌风险。FANCJ 编码一种 DNA 解旋酶,该酶参与双链断裂 (DSBs) 和链间交联 (ICLs) 的同源重组 (HR) 修复,但它的作用机制尚不清楚。在这里,我们通过活细胞成像显示,FANCJ 募集到激光诱导的 DSB 但不募集到补骨脂素诱导的 ICL,这依赖于具有核酸酶活性的 MRE11。FANCJ 与 MRE11 直接相互作用,并以特定的方式抑制其外切核酸酶活性,表明 FANCJ 调节 MRE11 核酸酶以促进 DSB 加工和适当的末端切除。缺乏 FANCJ 和 MRE11 的细胞显示出增加的电离辐射 (IR) 抗性、γH2AX 和 RAD51 焦点数量减少以及 DNA 依赖性蛋白激酶催化亚基焦点数量增加,表明 HR 受损并且非同源末端连接 (NHEJ) 途径被募集来帮助细胞应对 IR 诱导的链断裂。FANCJ 和 MRE11 之间的相互作用确保了对 IR 诱导的 DSB 的正常反应,而 FANCJ 参与 ICL 修复受到 MLH1 和 FA 途径的调节。我们的发现结合 HR 修复的当前模型进行了讨论。