Park C H, Mu D, Reardon J T, Sancar A
Department of Biochemistry and Biophysics, University of North Carolina School of Medicine, Chapel Hill 27599.
J Biol Chem. 1995 Mar 3;270(9):4896-902. doi: 10.1074/jbc.270.9.4896.
Recent studies have revealed that the general transcription factor TFIIH is also a general excision repair factor which, along with several other proteins, is required for transcription-independent excision reaction. As a general transcription factor, TFIIH is recruited to RNA polymerase II-promoter complex by another general transcription factor called TFIIE. We were interested in knowing whether TFIIE is also involved in recruiting TFIIH to the excision repair complex. We found that cell-free extract depleted of TFIIE carried out excision repair at a normal rate, leading us to conclude that TFIIE is not involved in recruiting TFIIH to the damage site and has no role in general excision repair. In contrast, the human damage recognition protein XPA specifically binds to TFIIH and apparently recruits it to the damage site. The carboxyl-terminal half of XPA is responsible for specific interaction with TFIIH. The C261S/C264S mutant of XPA bound the ERCC1-XPF complex normally, but failed to bind TFIIH and failed to complement an XP-A mutant cell-free extract indicating that the XPA-TFIIH interaction is essential to effecting the excision reaction. Interestingly, XPA also binds to the p34 subunit of TFIIE specifically and in competition with the p56 subunit of TFIIE. This latter interaction has no apparent role in general excision repair but may be relevant in the transcription-coupled repair reaction.
最近的研究表明,通用转录因子TFIIH也是一种通用切除修复因子,它与其他几种蛋白质一起,是转录非依赖性切除反应所必需的。作为一种通用转录因子,TFIIH被另一种称为TFIIE的通用转录因子招募到RNA聚合酶II启动子复合物中。我们想知道TFIIE是否也参与将TFIIH招募到切除修复复合物中。我们发现,去除TFIIE的无细胞提取物以正常速率进行切除修复,这使我们得出结论,TFIIE不参与将TFIIH招募到损伤位点,并且在通用切除修复中不起作用。相比之下,人类损伤识别蛋白XPA特异性结合TFIIH,并显然将其招募到损伤位点。XPA的羧基末端一半负责与TFIIH的特异性相互作用。XPA的C261S/C264S突变体正常结合ERCC1-XPF复合物,但未能结合TFIIH,也未能补充XP-A突变体无细胞提取物,这表明XPA-TFIIH相互作用对于实现切除反应至关重要。有趣的是,XPA还特异性结合TFIIE的p34亚基,并与TFIIE的p56亚基竞争。后一种相互作用在通用切除修复中没有明显作用,但可能与转录偶联修复反应有关。