title: Tripartite DNA Lesion Recognition and Verification by XPC, TFIIH, and XPA in Nucleotide Excision Repair.
abstract: Transcription factor IIH (TFIIH) is essential for both transcription and nucleotide excision repair (NER). DNA lesions are initially detected by NER factors XPC and XPE or stalled RNA polymerases, but only bulky lesions are preferentially repaired by NER. To elucidate substrate specificity in NER, we have prepared homogeneous human ten-subunit TFIIH and its seven-subunit core (Core7) without the CAK module and show that bulky lesions in DNA inhibit the ATPase and helicase activities of both XPB and XPD in Core7 to promote NER, whereas non-genuine NER substrates have no such effect. Moreover, the NER factor XPA activates unwinding of normal DNA by Core7, but inhibits the Core7 helicase activity in the presence of bulky lesions. Finally, the CAK module inhibits DNA binding by TFIIH and thereby enhances XPC-dependent specific recruitment of TFIIH. Our results support a tripartite lesion verification mechanism involving XPC, TFIIH, and XPA for efficient NER.
authors.affiliation: Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Chia-Lung
authors.initials: CL
authors.lastname: Li
authors.affiliation: Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Filip M
authors.initials: FM
authors.lastname: Golebiowski
authors.affiliation: Biosignal Research Center, Organization of Advanced Science and Technology, Kobe University, Kobe 657-8501, Japan; Department of Biology, Graduate School of Science, Kobe University, Kobe 657-8501, Japan.
authors.forename: Yuki
authors.initials: Y
authors.lastname: Onishi
authors.affiliation: Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA.
authors.forename: Nadine L
authors.initials: NL
authors.lastname: Samara
authors.affiliation: Biosignal Research Center, Organization of Advanced Science and Technology, Kobe University, Kobe 657-8501, Japan; Department of Biology, Graduate School of Science, Kobe University, Kobe 657-8501, Japan.
authors.forename: Kaoru
authors.initials: K
authors.lastname: Sugasawa
authors.affiliation: Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD 20892, USA. Electronic address: wei.yang@nih.gov.
authors.forename: Wei
authors.initials: W
authors.lastname: Yang
mesh_terms: D000818:Animals; D002945:Cisplatin / Q000737:chemistry; D018736:DNA Adducts / Q000737:chemistry / Q000235:genetics*; D004260:DNA Repair; D004277:DNA, Single-Stranded / Q000502:physiology; D004268:DNA-Binding Proteins / Q000737:chemistry / Q000502:physiology*; D024202:Electrophoretic Mobility Shift Assay; D006801:Humans; D011485:Protein Binding; D061987:Sf9 Cells; D018411:Spodoptera; D051758:Transcription Factor TFIIH / Q000737:chemistry / Q000502:physiology*; D051760:Xeroderma Pigmentosum Group A Protein / Q000737:chemistry / Q000502:physiology*
chemical_list: D018736:DNA Adducts; D004277:DNA, Single-Stranded; D004268:DNA-Binding Proteins; C497314:XPA protein, human; D051760:Xeroderma Pigmentosum Group A Protein; D051758:Transcription Factor TFIIH; C086984:XPC protein, human; D002945:Cisplatin
keywords: Cy5; Walker A motif; abasic site; cisplatin; motor; mutation