title: Dissociation of CAK from core TFIIH reveals a functional link between XP-G/CS and the TFIIH disassembly state.
abstract: Transcription factor II H (TFIIH) is comprised of core TFIIH and Cdk-activating kinase (CAK) complexes. Here, we investigated the molecular and cellular manifestation of the TFIIH compositional changes by XPG truncation mutations. We showed that both core TFIIH and CAK are rapidly recruited to damage sites in repair-proficient cells. Chromatin immunoprecipitation against TFIIH and CAK components revealed a physical engagement of CAK in nucleotide excision repair (NER). While XPD recruitment to DNA damage was normal, CAK was not recruited in severe XP-G and XP-G/CS cells, indicating that the associations of CAK and XPD to core TFIIH are differentially affected. A CAK inhibition approach showed that CAK activity is not required for assembling pre-incision machinery in vivo or for removing genomic photolesions. Instead, CAK is involved in Ser5-phosphorylation and UV-induced degradation of RNA polymerase II. The CAK inhibition impaired transcription from undamaged and UV-damaged reporter, and partially decreased transcription of p53-dependent genes. The overall results demonstrated that a) XP-G/CS mutations affect the disassembly state of TFIIH resulting in the dissociation of CAK, but not XPD from core TFIIH, and b) CAK activity is not essential for global genomic repair but involved in general transcription and damage-induced RNA polymerase II degradation.
authors.affiliation: Department of Radiology, The Ohio State University, Columbus, Ohio, USA.
authors.forename: Hany H
authors.initials: HH
authors.lastname: Arab
authors.forename: Gulzar
authors.initials: G
authors.lastname: Wani
authors.forename: Alo
authors.initials: A
authors.lastname: Ray
authors.forename: Zubair I
authors.initials: ZI
authors.lastname: Shah
authors.forename: Qianzheng
authors.initials: Q
authors.lastname: Zhu
authors.forename: Altaf A
authors.initials: AA
authors.lastname: Wani
mesh_terms: D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D018844:Cyclin-Dependent Kinases / Q000378:metabolism*; D004249:DNA Damage; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004720:Endonucleases / Q000378:metabolism*; D009687:Nuclear Proteins / Q000378:metabolism*; D010766:Phosphorylation; D051758:Transcription Factor TFIIH / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism*; D014466:Ultraviolet Rays; D000095043:Cyclin-Dependent Kinase-Activating Kinase; D000099250:DNA Excision Repair Protein ERCC-5
chemical_list: D018844:Cyclin-Dependent Kinases; D004268:DNA-Binding Proteins; D004720:Endonucleases; D009687:Nuclear Proteins; D051758:Transcription Factor TFIIH; D014157:Transcription Factors; D000095043:Cyclin-Dependent Kinase-Activating Kinase; D000099250:DNA Excision Repair Protein ERCC-5