PRMT5 dimethylates R30 of the p65 subunit to activate NF-κB

• DOI: 10.1073/pnas.1311784110
• Publication Date: 2013-08-01T04:46:56Z
• Authors: Han Wei; Benlian Wang; Masaru Miyagi; Yun She; Banu Gopalan; De-Bin Huang; Gourisankar Ghosh; George R. Stark; Tao Lu
• Keywords: 0301 basic medicine; 570; Protein-Arginine N-Methyltransferases; Blotting, Western; Oligonucleotides; Electrophoretic Mobility Shift Assay; histone; Methylation; Cell Line; 03 medical and health sciences; Tandem Mass Spectrometry; Cell Line, Tumor; Genetics; 2.1 Biological and endogenous factors; Humans; Immunoprecipitation; Aetiology; Luciferases; Protein Processing; Cancer; mass spectrometry; Tumor; Blotting; Post-Translational; Transcription Factor RelA; Microarray Analysis; Gene Expression Regulation; Western; Protein Processing, Post-Translational; Biotechnology

The ubiquitous inducible transcription factor NF-κB plays central roles in immune and inflammatory responses tumorigenesis. Complex posttranslational modifications of the p65 subunit (RelA) are a major aspect extremely flexible regulation activity. Although phosphorylation, acetylation, ubiquitination, lysine methylation have been well described, arginine has not yet found. We now report that, response to IL-1β, is dimethylated on 30 (R30) by protein-arginine methyltransferase 5 (PRMT5). Expression R30A R30K mutants substantially decreased ability bind κB elements drive gene expression. A model which dimethyl R30 placed into crystal structure predicts new van der Waals contacts that stabilize intraprotein interactions indirectly increase affinity for DNA. PRMT5 was only coprecipitated with p65, its overexpression increased activity, whereas knockdown had opposite effect. Microarray analysis revealed ∼85% NF-κB–inducible genes down-regulated mutation similarly knocking down. Many cytokine chemokine among these, conditioned media from cells expressing mutant much less NF-κB–inducing activity than wild-type protein. overexpressed many types cancer, often striking degree, indicating high levels this enzyme may promote tumorigenesis, at least part facilitating NF-κB-induced