Regulation of the bHLH transcription factor E2A in epithelial cells by interaction with the Na/K-ATPase beta1 subunit

Archives of Biochemistry and Biophysics
Matthew Plotkin, Lex Pelger

Abstract

The bHLH transcription factor E2A controls proliferation and differentiation in many cell types including kidney epithelial cells. To identify regulatory binding partners of E2A in the kidney, a yeast two-hybrid assay with a human adult kidney cDNA library was performed. Results demonstrated E2A interactions with other HLH proteins including Id1-3 and Pod1 and the Na/K-ATPase beta1 subunit. The specificity of beta1 subunit binding was confirmed by co-immunoprecipitation of E2A and beta1 subunit deletion constructs in HEK cells demonstrating E2A binding to the cytoplasmic tail of the beta1 subunit. Immunofluorescence and Western analysis of HEK cells co-transfected with GFP-beta1 subunit and E2A demonstrated E2A membrane binding and increased beta1 subunit membrane localization. Increased beta1 subunit expression resulted in decreased nuclear E2A expression and protein half-life and reduced E2A induced gene expression. These results suggest that E2A and Na/K-ATPase beta1 subunit expression in epithelial cells are regulated by interactions between these proteins.

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