Huber P A, Medhurst A L, Youssoufian H, Mathew C G
Division of Medical Genetics, Guy's, King's and St. Thomas' School of Medicine, Guy's Hospital, 7th Floor, Guy's Tower, London, SE1 9RT, United Kingdom.
Biochem Biophys Res Commun. 2000 Feb 5;268(1):73-7. doi: 10.1006/bbrc.1999.2055.
Fanconi anemia is a chromosomal breakage disorder with eight complementation groups (A-H), and three genes (FANCA, FANCC, and FANCG) have been identified. Initial investigations of the interaction between FANCA and FANCC, principally by co-immunoprecipitation, have proved controversial. We used the yeast two-hybrid assay to test for interactions of the FANCA, FANCC, and FANCG proteins. No activation of the reporter gene was observed in yeast co-expressing FANCA and FANCC as hybrid proteins, suggesting that FANCA does not directly interact with FANCC. However, a high level of activation was found when FANCA was co-expressed with FANCG, indicating strong, direct interaction between these proteins. Both FANCA and FANCG show weak but consistent interaction with themselves, suggesting that their function may involve dimerisation. The site of interaction of FANCG with FANCA was investigated by analysis of 12 mutant fragments of FANCG. Although both N- and C-terminal fragments did interact, binding to FANCA was drastically reduced, suggesting that more than one region of the FANCG protein is required for proper interaction with FANCA.
范可尼贫血是一种具有八个互补组（A - H）的染色体断裂疾病，并且已经鉴定出三个基因（FANCA、FANCC和FANCG）。最初对FANCA和FANCC之间相互作用的研究，主要通过免疫共沉淀法进行，结果存在争议。我们使用酵母双杂交试验来检测FANCA、FANCC和FANCG蛋白之间的相互作用。在共表达FANCA和FANCC作为杂交蛋白的酵母中未观察到报告基因的激活，这表明FANCA不直接与FANCC相互作用。然而，当FANCA与FANCG共表达时发现了高水平的激活，表明这些蛋白之间存在强烈的直接相互作用。FANCA和FANCG都显示出与自身的弱但一致的相互作用，这表明它们的功能可能涉及二聚化。通过分析FANCG的12个突变片段研究了FANCG与FANCA的相互作用位点。虽然N端和C端片段确实发生了相互作用，但与FANCA的结合大幅减少，这表明FANCG蛋白需要多个区域才能与FANCA进行适当的相互作用。
