title: Structural and functional characterization of interactions involving the Tfb1 subunit of TFIIH and the NER factor Rad2.
abstract: The general transcription factor IIH (TFIIH) plays crucial roles in transcription as part of the pre-initiation complex (PIC) and in DNA repair as part of the nucleotide excision repair (NER) machinery. During NER, TFIIH recruits the 3'-endonuclease Rad2 to damaged DNA. In this manuscript, we functionally and structurally characterized the interaction between the Tfb1 subunit of TFIIH and Rad2. We show that deletion of either the PH domain of Tfb1 (Tfb1PH) or several segments of the Rad2 spacer region yield yeast with enhanced sensitivity to UV irradiation. Isothermal titration calorimetry studies demonstrate that two acidic segments of the Rad2 spacer bind to Tfb1PH with nanomolar affinity. Structure determination of a Rad2-Tfb1PH complex indicates that Rad2 binds to TFIIH using a similar motif as TFIIEα uses to bind TFIIH in the PIC. Together, these results provide a mechanistic bridge between the role of TFIIH in transcription and DNA repair.
authors.affiliation: Département de Biochimie, Université de Montréal, C.P. 6128, Succursale Centre-Ville, Montréal, QC H3C 3J7, Canada.
authors.forename: Julien
authors.initials: J
authors.lastname: Lafrance-Vanasse
authors.forename: Geneviève
authors.initials: G
authors.lastname: Arseneault
authors.forename: Laurent
authors.initials: L
authors.lastname: Cappadocia
authors.forename: Hung-Ta
authors.initials: HT
authors.lastname: Chen
authors.forename: Pascale
authors.initials: P
authors.lastname: Legault
authors.forename: James G
authors.initials: JG
authors.lastname: Omichinski
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000378:metabolism; D004706:Endodeoxyribonucleases / Q000737:chemistry* / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019906:Nuclear Magnetic Resonance, Biomolecular; D017434:Protein Structure, Tertiary; D021122:Protein Subunits / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011836:Radiation Tolerance; D012441:Saccharomyces cerevisiae / Q000235:genetics / Q000254:growth & development / Q000528:radiation effects; D029701:Saccharomyces cerevisiae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D017384:Sequence Deletion; D019473:Transcription Factors, TFII / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry; D014466:Ultraviolet Rays
chemical_list: D004268:DNA-Binding Proteins; D021122:Protein Subunits; D029701:Saccharomyces cerevisiae Proteins; C086993:TFB1 protein, S cerevisiae; D019473:Transcription Factors, TFII; D016159:Tumor Suppressor Protein p53; C066644:transcription factor TFIIE; C073998:RAD2 protein, S cerevisiae; D004706:Endodeoxyribonucleases