Coin Frédéric, Auriol Jérome, Tapias Angel, Clivio Pascale, Vermeulen Wim, Egly Jean-Marc
Institut de Génétique et de Biologie Moléculaire et Cellulaire, CNRS/INSERM/ULP, Illkirch, CU Strasbourg, France.
EMBO J. 2004 Dec 8;23(24):4835-46. doi: 10.1038/sj.emboj.7600480. Epub 2004 Nov 18.
Nucleotide excision repair (NER) removes damage from DNA in a tightly regulated multiprotein process. The xeroderma pigmentosum group B (XPB) helicase subunit of TFIIH functions in NER and transcription. The serine 751 (S751) residue of XPB was found to be phosphorylated in vivo. This phosphorylation inhibits NER and the microinjection of a phosphomimicking XPB-S751E mutant is unable to correct the NER defect of XP-B cells. Conversely, XPB-S751 dephosphorylation or its substitution with alanine (S751A) restores NER both in vivo and in vitro. Surprisingly, phospho/dephosphorylation of S751 spares TFIIH-dependent transcription. Finally, the phosphorylation of XPB-S751 does not impair the TFIIH unwinding of the DNA around the lesion, but rather prevents the 5' incision triggered by the ERCC1-XPF endonuclease. These data support an additional role for XPB in promoting the incision of the damaged fragment and reveal a point of NER regulation on TFIIH without interference in its transcription activity.
核苷酸切除修复(NER)在一个严格调控的多蛋白过程中去除DNA损伤。TFIIH的着色性干皮病B组(XPB)解旋酶亚基在NER和转录过程中发挥作用。发现XPB的丝氨酸751(S751)残基在体内被磷酸化。这种磷酸化抑制NER,并且显微注射模拟磷酸化的XPB - S751E突变体无法纠正XP - B细胞的NER缺陷。相反,XPB - S751去磷酸化或用丙氨酸替代(S751A)在体内和体外均可恢复NER。令人惊讶的是,S751的磷酸化/去磷酸化并不影响TFIIH依赖的转录。最后,XPB - S751的磷酸化并不损害TFIIH对损伤周围DNA的解旋,而是阻止了由ERCC1 - XPF核酸内切酶触发的5'切口。这些数据支持了XPB在促进受损片段切口方面的额外作用,并揭示了NER在TFIIH上的一个调控点,而不会干扰其转录活性。