A conserved mechanism for binding of p53 DNA-Binding domain and Anti-Apoptotic Bcl-2 family proteins

• 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
• Other Authors: School of Biological Sciences
• Format: Article
• Language: English
• Published: 2018
• Subjects: DRNTU::Science::Biological sciences; Apoptosis; Bcl-2 Family Proteins
• Online Access: https://hdl.handle.net/10356/87108; http://hdl.handle.net/10220/46692
• Institution: Nanyang Technological University

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.