A requirement of FancL and FancD2 monoubiquitination in DNA repair
A requirement of FancL and FancD2 monoubiquitination in DNA repair
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DOI：
10.1111/j.1365-2443.2007.01054.x
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发表时间：
2007-03-01
期刊：
影响因子：
2.1
通讯作者：
Ishiai, Masamichi
中科院分区：
文献类型：
--
作者：
Seki, Sohsuke;Ohzeki, Mioko;Ishiai, Masamichi
罕见的遗传性疾病范可尼贫血（FA）可能是由FA核心复合物（FancA/B/C/E/F/G/L/M）、关键调节因子FancD 2、乳腺癌易感蛋白BRCA 2/FancD 1或新鉴定的FancJ/BRIP 1解旋酶的组分突变引起的。通过使用N-末端鸡（ch）FancD 2作为诱饵进行酵母双杂交（Y2 H）筛选，我们已经鉴定了chFancL，其可能是FA核心复合物的泛素E3连接酶亚基。我们还发现异位表达的FancD 2和FancL在293 T细胞中共免疫沉淀，并且这种相互作用依赖于FancL的PHD结构域。FANCL破坏的鸡DT 40细胞在FancD 2单泛素化和病灶形成方面都显示出缺陷。重要的是，缺乏FANCL或FANCD 2基因的细胞系，或携带FancD 2单泛素化位点（其中Lys 563残基变为Arg）的“敲入”突变的细胞系，在通过同源重组（HR）修复I-SceI诱导的染色体断裂中显示出数量上相同的缺陷。这些数据确立了FANCL和FancD 2单泛素化在HR修复中的作用。
The rare hereditary disorder Fanconi anemia (FA) can be caused by mutations in components of the FA core complex (FancA/B/C/E/F/G/L/M), a key regulator FancD2, the breast cancer susceptibility protein BRCA2/FancD1, or the newly identified FancJ/BRIP1 helicase. By performing yeast two-hybrid (Y2H) screens using N-terminal chicken (ch) FancD2 as a bait, we have identified chFancL, the likely ubiquitin E3 ligase subunit of the FA core complex. We also found that ectopically expressed FancD2 and FancL co-immunoprecipitated in 293T cells, and this interaction was dependent on the PHD domain of FancL. FANCL-disrupted chicken DT40 cells displayed defects in both FancD2 monoubiquitination and focus formation. Importantly, cell lines lacking the FANCL or FANCD2 genes, or carrying a "knock-in" mutation of the FancD2 monoubiquitination site (where the Lys 563 residue is changed to Arg), displayed quantitatively identical defects in the repair of I-SceI-induced chromosomal breaks by homologous recombination (HR). These data establish the role of FANCL and FancD2 monoubiquitination in HR repair.