title: A conserved mechanism for binding of p53 DNA-binding domain and anti-apoptotic Bcl-2 family proteins.
abstract: The molecular interaction between tumor suppressor p53 and the anti-apoptotic Bcl-2 family proteins plays an essential role in the transcription-independent apoptotic pathway of p53. In this study, we investigated the binding of p53 DNA-binding domain (p53DBD) with the anti-apoptotic Bcl-2 family proteins, Bcl-w, Mcl-1, and Bcl-2, using GST pull-down assay and NMR spectroscopy. The GST pull-down assays and NMR experiments demonstrated the direct binding of the p53DBD with Bcl-w, Mcl-1, and Bcl-2. Further, NMR chemical shift perturbation data showed that Bcl-w and Mcl-1 bind to the positively charged DNA-binding surface of p53DBD. Noticeably, the refined structural models of the complexes between p53DBD and Bcl-w, Mcl-1, and Bcl-2 showed that the binding mode of p53DBD is highly conserved among the anti-apoptotic Bcl-2 family proteins. Furthermore, the chemical shift perturbations on Bcl-w, Mcl-1, and Bcl-2 induced by p53DBD binding occurred not only at the p53DBD-binding acidic region but also at the BH3 peptide-binding pocket, which suggests an allosteric conformational change similar to that observed in Bcl-XL. Taken altogether, our results revealed a structural basis for a conserved binding mechanism between p53DBD and the anti-apoptotic Bcl-2 family proteins, which shed light on to the molecular understanding of the transcription-independent apoptosis pathway of p53.
authors.affiliation: Medical Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon 305-806, Korea.
authors.forename: Dong-Hwa
authors.initials: DH
authors.lastname: Lee
authors.forename: Ji-Hyang
authors.initials: JH
authors.lastname: Ha
authors.forename: Yul
authors.initials: Y
authors.lastname: Kim
authors.forename: Mi
authors.initials: M
authors.lastname: Jang
authors.forename: Sung Jean
authors.initials: SJ
authors.lastname: Park
authors.forename: Ho Sup
authors.initials: HS
authors.lastname: Yoon
authors.forename: Eun-Hee
authors.initials: EH
authors.lastname: Kim
authors.forename: Kwang-Hee
authors.initials: KH
authors.lastname: Bae
authors.forename: Byoung Chul
authors.initials: BC
authors.lastname: Park
authors.forename: Sung Goo
authors.initials: SG
authors.lastname: Park
authors.forename: Gwan-Su
authors.initials: GS
authors.lastname: Yi
authors.forename: Seung-Wook
authors.initials: SW
authors.lastname: Chi
mesh_terms: D000494:Allosteric Regulation; D051017:Apoptosis Regulatory Proteins / Q000737:chemistry / Q000502:physiology; D002460:Cell Line; D006801:Humans; D008958:Models, Molecular; D064549:Myeloid Cell Leukemia Sequence 1 Protein / Q000737:chemistry* / Q000502:physiology; D019906:Nuclear Magnetic Resonance, Biomolecular; D010449:Peptide Mapping; D011485:Protein Binding; D054730:Protein Interaction Domains and Motifs; D020836:Protein Structure, Quaternary; D017433:Protein Structure, Secondary; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000737:chemistry* / Q000502:physiology; D016159:Tumor Suppressor Protein p53 / Q000737:chemistry* / Q000502:physiology
chemical_list: D051017:Apoptosis Regulatory Proteins; C491073:BCL2L2 protein, human; C579095:MCL1 protein, human; D064549:Myeloid Cell Leukemia Sequence 1 Protein; D019253:Proto-Oncogene Proteins c-bcl-2; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53