DNA damage recognition by XPA protein promotes efficient recruitment of transcription factor II H.
The human basal transcription factor IIH (TFIIH) is an essential component of the nucleotide excision repair machinery. TFIIH is required for reaction steps concomitant with or prior to the formation of dual incisions in the damaged DNA strand. To understand the mechanism underlying the recruitment of TFIIH to DNA damage sites we have analyzed i) the direct affinity of TFIIH for damaged or undamaged DNA and ii) the interaction of TFIIH with XPA.DNA complexes, formed using unirradiated or UV-irradiated DNA. Filter binding assays showed that TFIIH has some affinity for the DNA, but in contrast to XPA, does not show any preference for UV-irradiated DNA. Pull-down experiments demonstrated that TFIIH binds to XPA.DNA complexes in an UV damage-dependent manner by a direct protein-protein interaction with XPA. We propose that an enhancement of the affinity of XPA protein for TFIIH could arise from conformational changes of XPA when it binds to UV lesions on the DNA.
Nocentini
S
S
CNRS UMR 218 et LRC no. 1 du CEA, Institut Curie, Section de Recherche, 26 rue d'Ulm, 75248 Paris Cedex 05, France.
Coin
F
F
Saijo
M
M
Tanaka
K
K
Egly
J M
JM
D004247:DNA / Q000378:metabolism / Q000528:radiation effects; D004249:DNA Damage; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism*; D008957:Models, Genetic; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D051716:Replication Protein A; D051758:Transcription Factor TFIIH; D014157:Transcription Factors / Q000378:metabolism*; D019473:Transcription Factors, TFII; D014466:Ultraviolet Rays; D051760:Xeroderma Pigmentosum Group A Protein
D004268:DNA-Binding Proteins; D051716:Replication Protein A; D014157:Transcription Factors; D019473:Transcription Factors, TFII; D051760:Xeroderma Pigmentosum Group A Protein; D051758:Transcription Factor TFIIH; D004247:DNA