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  • Sam68 functions as a transcriptional coactivator of the p53 tumor suppressor.

Sam68 functions as a transcriptional coactivator of the p53 tumor suppressor.

Nucleic acids research (2016-07-02)
Naomi Li, Stéphane Richard
ABSTRACT

Sam68 is a known sequence-specific RNA binding protein that regulates alternative splicing events during the cell cycle and apoptosis. Sam68 has also been shown to influence transcription, but the molecular mechanism remains undefined. Herein we identify Sam68 as a transcriptional coactivator of the p53 tumor suppressor in response to DNA damage. Using CRISPR/Cas9 generated isogenic HCT116 Sam68-/- cell lines wild type or deficient for p53, we show that Sam68 is required for the efficient transactivation of p53 target genes. Consistently, Sam68 depletion caused defects in DNA damage-induced cell cycle arrest and apoptosis mediated by p53. Mechanistically, we demonstrate that Sam68 physically interacted with p53 in an RNA-dependent manner, and that this interaction was essential for the coactivator function of Sam68. Furthermore, we show that both Sam68 and p53 were recruited to promoters of p53-responsive genes, suggesting interdependence. Finally, Sam68 acted in concert with the p53 long noncoding RNA (lncRNA) target PR-lncRNA-1 for p53 recruitment, implicating a positive-feedback mechanism in which lncRNAs induced by the Sam68/p53 complex can enhance p53 transcriptional activity. These findings define a hitherto novel mechanism of action for Sam68 in governing p53 transcriptional activation, and represent the first report of Sam68 in the regulation of tumor suppressor activities.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-Sam 68 Antibody, serum, Upstate®
Sigma-Aldrich
Anti-Actin antibody, Mouse monoclonal, clone AC-40, purified from hybridoma cell culture
Sigma-Aldrich
Anti-Myc Tag Antibody, clone 4A6, clone 4A6, Upstate®, from mouse