APE1/Ref-1 enhances DNA binding activity of mutant p53 in a redox-dependent manner

Cun,Yanping; Dai,Nan; Li,Mengxia; Xiong,Chengjie; Zhang,Qinhong; Sui,Jiangdong; Qian,Chengyuan; Wang,Dong

doi:10.3892/or.2013.2892

APE1/Ref-1 enhances DNA binding activity of mutant p53 in a redox-dependent manner

Authors:
- Yanping Cun
- Nan Dai
- Mengxia Li
- Chengjie Xiong
- Qinhong Zhang
- Jiangdong Sui
- Chengyuan Qian
- Dong Wang
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Affiliations: Cancer Center, Daping Hospital and Research Institute of Surgery, Third Military Medical University, Chongqing, P.R. China, Department of Orthopedics, Wuhan General Hospital, Guangzhou Military Area Command, Wuhan, P.R. China
Published online on: December 2, 2013 https://doi.org/10.3892/or.2013.2892
Pages: 901-909

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Abstract

Apurinic/apyrimidinic endonuclease 1/redox factor-1 (APE1/Ref-1) is a dual function protein; in addition to its DNA repair activity, it can stimulate DNA binding activity of numerous transcription factors as a reduction-oxidation (redox) factor. APE1/Ref-1 has been found to be a potent activator of wild-type p53 (wtp53) DNA binding in vitro and in vivo. Although p53 is mutated in most types of human cancer including hepatocellular carcinoma (HCC), little is known about whether APE1/Ref-1 can regulate mutant p53 (mutp53). Herein, we reported the increased APE1/Ref-1 protein and accumulation of mutp53 in HCC by immunohistochemistry. Of note, it was observed that APE1/Ref-1 high-expression and mutp53 expression were associated with carcinogenesis and progression of HCC. To determine whether APE1/Ref-1 regulates DNA binding of mutp53, we performed electromobility shift assays (EMSAs) and quantitative chromatin immunoprecipitation (ChIP) assays in HCC cell lines. In contrast to sequence-specific and DNA structure-dependent binding of wtp53, reduced mutp53 efficiently bound to nonlinear DNA, but not to linear DNA. Notably, overexpression of APE1/Ref-1 resulted in increased DNA binding activity of mutp53, while downregulation of APE1/Ref-1 caused a marked decrease of mutp53 DNA binding. In addition, APE1/Ref-1 could not potentiate the accumulation of p21 mRNA and protein in mutp53 cells. These data indicate that APE1/Ref-1 can stimulate mutp53 DNA binding in a redox-dependent manner.

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