A conserved mechanism for binding of p53 DNA-binding domain and anti-apoptotic Bcl-2 family proteins

• Published in: Molecules and cells 2014, 37(3), , pp.264-269
• Main Authors: Lee, Dong-Hwa, Ha, Ji-Hyang, Kim, Yul, Jang, Mi, Park, Sung Jean, Yoon, Ho Sup, Kim, Eun-Hee, Bae, Kwang-Hee, Park, Byoung Chul, Park, Sung Goo, Yi, Gwan-Su, Chi, Seung-Wook
• Format: Article
• Language: English
• Subjects: Allosteric Regulation; Apoptosis Regulatory Proteins - chemistry; Apoptosis Regulatory Proteins - physiology; Cell Line; Humans; Models, Molecular; Myeloid Cell Leukemia Sequence 1 Protein - chemistry; Myeloid Cell Leukemia Sequence 1 Protein - physiology; Nuclear Magnetic Resonance, Biomolecular; Peptide Mapping; Protein Binding; Protein Interaction Domains and Motifs; Protein Structure, Quaternary; Protein Structure, Secondary; Proto-Oncogene Proteins c-bcl-2 - chemistry; Proto-Oncogene Proteins c-bcl-2 - physiology; Tumor Suppressor Protein p53 - chemistry; Tumor Suppressor Protein p53 - physiology; 생물학
• ISSN: 1016-8478; 0219-1032
• DOI: 10.14348/molcells.2014.0001

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.