p53 Requires an Intact C-Terminal Domain for DNA Binding and Transactivation

JOURNAL OF MOLECULAR BIOLOGY

卷:415

Article

Kim, Hyunjung³  Kim, Kyunghwan³  Choi, Jongkyu³  Heo, Kyu^3,4  Baek, Hwa Jin^1,2  Roeder, Robert G.¹  An, Woojin³ 

[1] Rockefeller Univ, Lab Biochem & Mol Biol, New York, NY 10065 USA

[2] Baylor Coll Med, Dept Biochem & Mol Biol, Houston, TX 77030 USA

[3] Univ So Calif, Norris Comprehens Canc Ctr, Dept Biochem & Mol Biol, Los Angeles, CA 90033 USA

[4] Dongnam Inst Radiol & Med Sci, Res Ctr, Pusan 619753, South Korea

关键词: p53;    C-terminal domain;    transcription;    DNA binding;    activation;   

DOI  :  10.1016/j.jmb.2011.12.001

来源: Elsevier

【 摘 要 】

The tumor suppressor p53 plays a critical role in mediating cellular response to a wide range of environmental stresses. p53 regulates these processes mainly by acting as a short-lived DNA binding protein that stimulates transcription from numerous genes involved in cell cycle arrest, programmed cell death, and other processes. To investigate the importance of the C-terminal domain of p53, we generated a series of deletion and point mutations in this region and analyzed their effects on p53 transcription activity. Our results show that C-terminal deletion and point mutations at K320 and K382 abolish p53-mediated transcription in the context of DNA or chromatin. This defect is specific for DNA molecules because inactive mutants fail to bind a consensus p53 response element in both free DNA and nucleosomes. Chromatin immunoprecipitation assays further substantiate the importance of the p53 C-terminal domain for the targeted localization of p53 and the concomitant recruitment of p300 onto p53-responsive genes. Moreover, a synthetic peptide comprising the last 30 amino acids of p53 interacts with the N-terminal and C-terminal domains of p53 and antagonizes p53-dependent transcription. Taken together, our data reveal a functional requirement for the p53 C-terminal domain in p53 transactivation and support a working model in which the C-terminus serves as a positive regulator for N-terminal activation and central DNA binding domains. (C) 2011 Elsevier Ltd. All rights reserved.

【 授权许可】

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