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Coordinate regulation by transcription factors and DNA methylation in the core promoter region of SIRT6 in bovine adipocytes

Author:
Jie-yun Hong, Chu-gang Mei, Shi-jun Li, Hong-bao Wang, Chun-ping Zhao, Lin-sen Zan
Source:
Archives of biochemistry and biophysics 2018 v.659 pp. 1-12
ISSN:
0003-9861
Subject:
DNA methylation, adipocytes, binding sites, cattle, gel electrophoresis, gene expression, glucose, insulin secretion, lipid metabolism, longissimus muscle, luciferase, messenger RNA, moieties, neoplasms, promoter regions, proto-oncogenes, subcutaneous fat, testes, transactivators, transcription (genetics), transferases
Abstract:
Sirtuin6 (SIRT6) is an ADP-ribosyltransferase and NAD⁺-dependent deacylase of acetyl groups and long-chain fatty acyl groups, and has been shown as a regulator of insulin secretion, glucose metabolism, lipid metabolism, and cancer. In this study, we determined that the bovine SIRT6 showed higher levels of mRNA expression in the testis, longissimus thoracis, and subcutaneous fat tissue. To elucidate the molecular regulation mechanism of bovine SIRT6 expression, we obtained a 2-kb fragment containing the 5′-regulatory region, and the functional proximal minimal promoter of bovine SIRT6 was identified in the −472/−73 bp region. The CCAAT enhancer binding protein beta (CEBPβ), paired box 6 (PAX6), Kruppel-like factor 2 (KLF2), myb proto-oncogene protein (CMYB), nuclear respiratory factor 1 (NRF1), and E2F transcription factor 1 (E2F1) binding sites, as transcriptional activators or repressors in the core promoter region of SIRT6, were determined by electrophoretic mobility shift assay (EMSA) experiments and luciferase reporter assays. In addition, the results from methylation assay and luciferase report assay showed that the bovine SIRT6 promoter activity was coordinately regulated by methylation and NRF1 or E2F1 during bovine adipocyte differentiation. Taken together, this study illuminated the underlying mechanism of methylation and transcription regulation of SIRT6 expression in bovine adipocytes.
Agid:
6153474