Transcription elongation factor Brd4-P-TEFb accelerates intestinal differentiation-associated SLC2A5 gene expression

Abstract

Expression of the fructose transporter gene SLC2A5 and histone acetylation in the transcribed region are induced by differentiation associated-signals such as glucocorticoids and p44/42 mitogen-activated protein kinase (MAPK) inhibition in small intestinal Caco-2 cells. We co-treated with glucocorticoid receptor agonist dexamethasone (Dex) and p44/42 MAPK inhibitor PD98059 (PD) in Caco-2 cells with or without Brd4 small hairpin (sh) RNA expression vector, and the cells were analyzed by qRT-PCR and chromatin immunoprecipitation assays. The small intestine of wild-type mice and Brd4 +/− mice during weaning period were analyzed by qRT-PCR. Co-treatment with Dex and PD increased binding of the bromodomain-containing protein-4 (Brd4)–positive transcriptional elongation factor-b (P-TEFb)–RNA polymerase II complex to acetylated histones in the transcribed region of SLC2A5 . Brd4-protein depletion by shRNA revealed that the association of these proteins on the transcribed region of SLC2A5 promoted gene expression in a Brd4-dependent manner. Expression of small-intestine Slc2a5 , but not another intestinal gene sucrase-isomaltase, during weaning period, was significantly lower in Brd4 +/− mice compared with wild-type mice. Brd4-P-TEFb plays a crucial role in differentiation-associated transcription of SLC2A5 gene in intestinal Caco-2 cells and in the small intestine of mice during weaning period. Histone acetylation and the transcription elongation factor Brd4 are important for SLC2A5 expression in the small intestine. • Brd4 regulates expression of SLC2A5 in human intestinal Caco-2 cells. • Brd4–P-TEFb and acetylated histones bind transcribed region of SLC2A5. • Brd4 regulates jejunal Slc2a5 expression during weaning period of mice.