title: p300/CBP acetyl transferases interact with and acetylate the nucleotide excision repair factor XPG.
abstract: Nucleotide excision repair (NER) is an important DNA repair mechanism through which cells remove bulky DNA lesions. Following DNA damage, the histone acetyltransferase (HAT) p300 (also referred to as lysine acetyltransferase or KAT) is known to associate with proliferating cell nuclear antigen (PCNA), a master regulator of DNA replication and repair processes. This interaction, which results in HAT inhibition, may be dissociated by the cell cycle inhibitor p21(CDKN1A), thereby restoring p300 activity; however, the role of this protein interplay is still unclear. Here, we report that silencing p300 or its homolog CREB-binding protein (CBP) by RNA interference (RNAi) significantly reduces DNA repair synthesis in human fibroblasts. In addition, we determined whether p300 and CBP may associate with and acetylate specific NER factors such as XPG, the 3'-endonuclease that is involved in the incision/excision step and is known to interact with PCNA. Our results show that p300 and CBP interact with XPG, which has been found to be acetylated in vivo. XPG is acetylated by p300 in vitro, and this reaction is inhibited by PCNA. Knocking down both p300/CBP by RNAi or by chemical inhibition with curcumin greatly reduced XPG acetylation, and a concomitant accumulation of the protein at DNA damage sites was observed. The ability of p21 to bind PCNA was found to regulate the interaction between p300 and XPG, and an abnormal accumulation of XPG at DNA damage sites was also found in p21(-/-) fibroblasts. These results indicate an additional function of p300/CBP in NER through the acetylation of XPG protein in a PCNA-p21 dependent manner.
authors.affiliation: Istituto di Genetica Molecolare (IGM) del CNR, Pavia, Italy.
authors.forename: Micol
authors.initials: M
authors.lastname: Tillhon
authors.forename: Ornella
authors.initials: O
authors.lastname: Cazzalini
authors.forename: Tiziana
authors.initials: T
authors.lastname: Nardo
authors.forename: Daniela
authors.initials: D
authors.lastname: Necchi
authors.forename: Sabrina
authors.initials: S
authors.lastname: Sommatis
authors.forename: Lucia A
authors.initials: LA
authors.lastname: Stivala
authors.forename: A Ivana
authors.initials: AI
authors.lastname: Scovassi
authors.forename: Ennio
authors.initials: E
authors.lastname: Prosperi
mesh_terms: D000107:Acetylation; D003474:Curcumin / Q000494:pharmacology; D050759:Cyclin-Dependent Kinase Inhibitor p21 / Q000378:metabolism; D053843:DNA Mismatch Repair; D004260:DNA Repair; D004268:DNA-Binding Proteins / Q000378:metabolism*; D004720:Endonucleases / Q000378:metabolism*; D005347:Fibroblasts / Q000201:enzymology; D006367:HeLa Cells; D006801:Humans; D009687:Nuclear Proteins / Q000378:metabolism*; D018809:Proliferating Cell Nuclear Antigen / Q000378:metabolism; D034741:RNA, Small Interfering; D014157:Transcription Factors / Q000378:metabolism*; D050880:p300-CBP Transcription Factors / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D000099250:DNA Excision Repair Protein ERCC-5
chemical_list: D003474:Curcumin; D050759:Cyclin-Dependent Kinase Inhibitor p21; D004268:DNA-Binding Proteins; D004720:Endonucleases; D009687:Nuclear Proteins; D018809:Proliferating Cell Nuclear Antigen; D034741:RNA, Small Interfering; D014157:Transcription Factors; D050880:p300-CBP Transcription Factors; D000099250:DNA Excision Repair Protein ERCC-5