Laboratory of Genome Maintenance, Rockefeller University, New York, NY 10065-6399, USA.
Blood. 2013 Jan 3;121(1):54-63. doi: 10.1182/blood-2012-07-441212. Epub 2012 Oct 23.
SLX4, the newly identified Fanconi anemia protein, FANCP, is implicated in repairing DNA damage induced by DNA interstrand cross-linking (ICL) agents, topoisomerase I (TOP1) inhibitors, and in Holliday junction resolution. It interacts with and enhances the activity of XPF-ERCC1, MUS81-EME1, and SLX1 nucleases, but the requirement for the specific nucleases in SLX4 function is unclear. Here, by complementing a null FA-P Fanconi anemia cell line with SLX4 mutants that specifically lack the interaction with each of the nucleases, we show that the SLX4-dependent XPF-ERCC1 activity is essential for ICL repair but is dispensable for repairing TOP1 inhibitor-induced DNA lesions. Conversely, MUS81-SLX4 interaction is critical for resistance to TOP1 inhibitors but is less important for ICL repair. Mutation of SLX4 that abrogates interaction with SLX1 results in partial resistance to both cross-linking agents and TOP1 inhibitors. These results demonstrate that SLX4 modulates multiple DNA repair pathways by regulating appropriate nucleases.
SLX4,一种新发现的范可尼贫血症蛋白,FANCP,与 DNA 链间交联(ICL)试剂、拓扑异构酶 I(TOP1)抑制剂诱导的 DNA 损伤修复有关,并参与 Holliday 连接的解决。它与 XPF-ERCC1、MUS81-EME1 和 SLX1 核酸酶相互作用并增强其活性,但 SLX4 功能中对特定核酸酶的需求尚不清楚。在这里,通过用 SLX4 突变体互补一个缺失的 FA-P 范可尼贫血细胞系,该突变体特异性缺乏与每种核酸酶的相互作用,我们表明 SLX4 依赖性 XPF-ERCC1 活性对于 ICL 修复是必需的,但对于修复 TOP1 抑制剂诱导的 DNA 损伤是可有可无的。相反,MUS81-SLX4 相互作用对于 TOP1 抑制剂的耐药性至关重要,但对于 ICL 修复的重要性较低。SLX4 突变导致与 SLX1 相互作用的缺失,导致对交联剂和 TOP1 抑制剂的部分耐药性。这些结果表明,SLX4 通过调节适当的核酸酶来调节多种 DNA 修复途径。