Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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A Conserved Mechanism for Binding of p53 DNA-Binding Domain and Anti-Apoptotic Bcl-2 Family Proteins

  • Lee, Dong-Hwa (Medical Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Ha, Ji-Hyang (Medical Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kim, Yul (Department of Bio and Brain Engineering, Korea Advanced Institute for Science and Technology) ;
  • Jang, Mi (Medical Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Sung Jean (College of Pharmacy, Gachon University) ;
  • Yoon, Ho Sup (Division of Structural Biology and Biochemistry, School of Biological Sciences, Nanyang Technological University) ;
  • Kim, Eun-Hee (Division of Magnetic Resonance, Korea Basic Science Institute) ;
  • Bae, Kwang-Hee (Research Center for Integrated Cellulomics, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Byoung Chul (Medical Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Park, Sung Goo (Medical Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Yi, Gwan-Su (Department of Bio and Brain Engineering, Korea Advanced Institute for Science and Technology) ;
  • Chi, Seung-Wook (Medical Proteomics Research Center, Korea Research Institute of Bioscience and Biotechnology)
  • Received : 2014.01.03
  • Accepted : 2014.01.14
  • Published : 2014.03.31
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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-$X_L$. 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.

Keywords

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