The ETS family transcription factor ELK-1 regulates induction of the cell cycle-regulatory gene p21 Waf1/Cip1and the BAX gene in sodium arsenite-exposed human keratinocyte HaCaT cells

Abstract

Cyclin-dependent kinase inhibitor (CDKN1A), often referred to as p21 Waf1/Cip1 (p21), is induced by a variety of environmental stresses. Transcription factor ELK-1 is a member of the ETS oncogene superfamily. Here, we show that ELK-1 directly transactivates the p21 gene, independently of p53 and EGR-1, in sodium arsenite (NaASO 2)-exposed HaCaT cells. Promoter deletion analysis and site-directed mutagenesis identified the presence of an ELK-1-binding core motif between -190 and -170 bp of the p21 promoter that confers inducibility by NaASO2. Chromatin immunoprecipitation and electrophoretic mobility shift analyses confirmed the specific binding of ELK-1 to its putative binding sequence within the p21 promoter. In addition, NaASO 2-induced p21 promoter activity was enhanced by exogenous expression of ELK-1 and reduced by expression of siRNA targeted to ELK-1 mRNA. The importance of ELK-1 in response to NaASO 2 was further confirmed by the observation that stable expression of ELK-1 siRNA in HaCaT cells resulted in the attenuation of NaASO 2-induced p21 expression. Although ELK-1 was activated by ERK, JNK, and p38 MAPK in response to NaASO 2, ELK-1-mediated activation of the p21 promoter was largely dependent on ERK. In addition, EGR-1 induced by ELK-1 seemed to be involved in NaASO 2-induced expression of BAX. This supports the view that the ERK/ELK-1 cascade is involved in p53-independent induction of p21 and BAX gene expression. © 2011 by The American Society for Biochemistry and Molecular Biology, Inc.