Direct interaction of the Fanconi anaemia protein FANCG with BRCA2/FANCD1
Direct interaction of the Fanconi anaemia protein FANCG with BRCA2/FANCD1
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DOI：
10.1093/hmg/ddg266
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发表时间：
2003-10-01
影响因子：
3.5
通讯作者：
Mathew, CG
中科院分区：
文献类型：
--
作者：
Hussain, S;Witt, E;Mathew, CG
范可尼贫血（FA）是一种常染色体隐性遗传疾病，以进行性骨髓衰竭、多种先天性异常和癌症风险增加为特征。FA细胞的特点是染色体不稳定和对DNA链间交联剂过敏。至少存在8个互补基团（FA-A到G），除FA-B外，其他所有互补基团的基因都已克隆。FANCA和FANCD2蛋白与BRCA1的相互作用，以及FANCD1基因与BRCA2相同的发现，证明了FA通路与乳腺癌易感性基因之间的功能联系。在这里，我们使用酵母双杂交系统来测试BRCA2或其效应体RAD51与FANCA、FANCC和FANCG蛋白之间的直接相互作用。我们发现FANCG能够结合BRCA2蛋白中的两个独立位点，位于BRC重复序列的两侧。此外，FANCG可以与人类细胞中的BRCA2共同免疫沉淀，FANCG可以在丝裂霉素c损伤DNA后与BRCA2和RAD51共同定位在核中心。这些结果表明，BRCA2直接与缺乏链间交联修复的途径相连，并且至少有一种FA蛋白与同源重组DNA修复机制密切相关。
Fanconi anaemia (FA) is an autosomal recessive genetic disorder characterized by progressive bone marrow failure, multiple congenital abnormalities, and an increased risk of cancer. FA cells are characterized by chromosomal instability and hypersensitivity to DNA interstrand crosslinking agents. At least eight complementation groups exist (FA-A to G), and the genes for all of these except FA-B have been cloned. Functional linkage between the FA pathway and genes involved in susceptibility to breast cancer has been demonstrated by the interaction of the FANCA and FANCD2 proteins with BRCA1, and the discovery that the FANCD1 gene is identical to BRCA2. Here we have used the yeast two-hybrid system to test for direct interaction between BRCA2 or its effector RAD51 and the FANCA, FANCC and FANCG proteins. We found that FANCG was capable of binding to two separate sites in the BRCA2 protein, located either side of the BRC repeats. Furthermore, FANCG could be co-immunoprecipitated with BRCA2 from human cells, and FANCG co-localized in nuclear foci with both BRCA2 and RAD51 following DNA damage with mitomycin C. These results demonstrate that BRCA2 is directly connected to a pathway that is deficient in interstrand crosslink repair, and that at least one other FA protein is closely associated with the homologous recombination DNA repair machinery.