title: p300 cooperates with c-Jun and PARP-1 at the p300 binding site to activate RhoB transcription in NSC126188-mediated apoptosis.
abstract: The anti-cancer agent NSC126188 induces apoptosis of stomach carcinoma NUGC-3 cells by inducing RhoB expression. Here, we present that the p300 binding site in the RhoB promoter is crucial for the binding of p300 and its partner transcription factors to activate RhoB transcription in NSC126188-mediated apoptosis. NSC126188 increased expression of p300 and c-Jun. Conversely, knockdown of p300 decreased RhoB expression in the presence of NSC126188. We found that poly(ADP-ribose) polymerase-1 (PARP-1) was associated with the p300 binding site and that PARP-1 knockdown inhibited NSC126188-mediated RhoB expression. In the cells treated with NSC126188, p300, PARP-1, and c-Jun interacted and bound the p300 binding site. Furthermore, chromatin immunoprecipitation (ChIP) analysis revealed strong p300 binding and weak c-Jun binding at the p300 binding site of RhoB promoter in cells treated with NSC126188. We also demonstrated that c-Jun played a crucial role in p300 binding. However, PARP-1 did not directly bind the p300 binding site, suggesting a bridging role between p300 and c-Jun. Electrophoretic mobility shift assays demonstrated a complex comprising p300/c-Jun/PARP-1 that bound wild type, but not a mutated, p300 binding site. In addition, overexpression of p300, PARP-1, or c-Jun dramatically enhanced RhoB promoter activity when it contained the wild type sequence but not mutated sequences, indicating the crucial role of the p300 binding site in NSC126188-induced transcription of RhoB. Taken together, these data suggest that p300 is recruited and cooperates with c-Jun and PARP-1 at the p300 binding site to activate RhoB transcription during NSC126188-mediated apoptosis.
authors.affiliation: BioMedical Genomics Research Center, KRIBB, Daejeon 305-806, Korea.
authors.forename: Bo-Kyung
authors.initials: BK
authors.lastname: Kim
authors.affiliation: BioMedical Genomics Research Center, KRIBB, Daejeon 305-806, Korea.
authors.forename: Joo-Young
authors.initials: JY
authors.lastname: Im
authors.affiliation: Dept. of Biotechnology, Yonsei University, Seoul 129-749, Korea.
authors.forename: Gyoonhee
authors.initials: G
authors.lastname: Han
authors.affiliation: BioMedical Genomics Research Center, KRIBB, Daejeon 305-806, Korea.
authors.forename: Woo-Jung
authors.initials: WJ
authors.lastname: Lee
authors.affiliation: BioMedical Genomics Research Center, KRIBB, Daejeon 305-806, Korea; Functional Genomics, University of Science and Technology, Daejeon 305-350, Korea.
authors.forename: Kyoung-Jae
authors.initials: KJ
authors.lastname: Won
authors.affiliation: BioMedical Genomics Research Center, KRIBB, Daejeon 305-806, Korea.
authors.forename: Kyung-Sook
authors.initials: KS
authors.lastname: Chung
authors.affiliation: School of Pharmacy, Dongguk University-Seoul, Seoul 100-715, Korea.
authors.forename: Kyeong
authors.initials: K
authors.lastname: Lee
authors.affiliation: BioMedical Genomics Research Center, KRIBB, Daejeon 305-806, Korea.
authors.forename: Hyun Seung
authors.initials: HS
authors.lastname: Ban
authors.affiliation: Dept. of Food Science and Technology, Chungnam National University, Daejeon 305-764, Korea.
authors.forename: KyungBin
authors.initials: K
authors.lastname: Song
authors.affiliation: BioMedical Genomics Research Center, KRIBB, Daejeon 305-806, Korea; Functional Genomics, University of Science and Technology, Daejeon 305-350, Korea. Electronic address: misun@kribb.re.kr.
authors.forename: Misun
authors.initials: M
authors.lastname: Won
mesh_terms: D017209:Apoptosis / Q000187:drug effects* / Q000235:genetics; D001665:Binding Sites; D045744:Cell Line, Tumor; D050881:E1A-Associated p300 Protein / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D010879:Piperazines / Q000494:pharmacology*; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases / Q000235:genetics / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D016755:Proto-Oncogene Proteins c-jun / Q000235:genetics / Q000378:metabolism*; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects*; D020743:rhoB GTP-Binding Protein / Q000235:genetics / Q000378:metabolism*
chemical_list: C558305:4-hexadecanoyl-1,1-dimethylpiperazin-1-ium; D010879:Piperazines; D016755:Proto-Oncogene Proteins c-jun; D014157:Transcription Factors; D050881:E1A-Associated p300 Protein; C494360:EP300 protein, human; C499320:PARP1 protein, human; D000071137:Poly (ADP-Ribose) Polymerase-1; D011065:Poly(ADP-ribose) Polymerases; D020743:rhoB GTP-Binding Protein
keywords: Apoptosis; NSC126188; PARP-1; RhoB; c-Jun; p300