The nuclear cofactor RAC3/AIB1/SRC-3 enhances Nrf2 signaling by interacting with transactivation domains

• Published in: Oncogene Vol. 32; no. 4; pp. 514 - 527
• Main Authors: Kim, J-H, Yu, S, Chen, J D, Kong, A N
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 24-01-2013; Nature Publishing Group
• Subjects: 631/45/612/822; 631/67; 631/80/86; Antioxidants; Apoptosis; Breast cancer; Cancer; Carcinogens; Cell Biology; Cell Line, Tumor; Development and progression; Gene expression; Genetic aspects; Health aspects; HeLa Cells; Heme Oxygenase-1 - genetics; Heme Oxygenase-1 - metabolism; Human Genetics; Humans; Internal Medicine; MCF-7 Cells; Medicine; Medicine & Public Health; NF-E2-Related Factor 2 - genetics; NF-E2-Related Factor 2 - metabolism; Nuclear Proteins - genetics; Nuclear Proteins - metabolism; Nuclear Receptor Coactivator 3 - genetics; Nuclear Receptor Coactivator 3 - metabolism; Oncology; original-article; Oxidases; Oxidative stress; Physiological aspects; Protein Binding; Protein Interaction Domains and Motifs; Proteins; rac GTP-Binding Proteins - genetics; rac GTP-Binding Proteins - metabolism; Signal Transduction; Trans-Activators - genetics; Trans-Activators - metabolism; Transcription Factors - genetics; Transcription Factors - metabolism; Transcriptional Activation; United States; RAC3/SRC-3; Nrf2; HO-1
• ISSN: 0950-9232; 1476-5594
• DOI: 10.1038/onc.2012.59

Nuclear factor erythroid 2-related factor 2 (Nrf2, NM 006164, 605 AA) is essential for the antioxidant responsive element (ARE)-mediated expression of a group of detoxifying antioxidant genes that detoxify carcinogens and protect against oxidative stress. Several proteins have been identified as Nrf2-interacting molecules. In this study, we found that the overexpression of receptor-associated coactivator 3 (RAC3)/AIB-1/steroid receptor coactivator-3, a nuclear coregulator and oncogene frequently amplified in human breast cancers, induced heme oxygenase-1 (HO-1) through Nrf2 transactivation in HeLa cells. Next, we determined the interaction between RAC3 and Nrf2 proteins using a co-immunoprecipitation assay and fluorescence resonance energy transfer analysis. The results showed that RAC3 bound directly to the Nrf2 protein in the nucleus. Subsequently, we identified the interacting domains of Nrf2 and RAC3 using a glutathione S-transferase pull-down assay. The results showed that both the N-terminal RAC3-pasB and C-terminal RAC3-R3B3 domains were tightly bound to the Neh4 and Neh5 transactivation domains. Furthermore, chromatin immunoprecipitation showed that RAC3 bound tightly to the ARE enhancer region of the HO-1 promoter via Nrf2 binding. These data suggest that Nrf2 activation is modulated and directly controlled through interactions with the RAC3 protein in HeLa cells.