Bioinformatics Analysis and Competitive Regulation by Transcription Factors of SIRT5 at the Core Promoter Region Using Bovine Adipocytes

Sirtuin 5 (SIRT5) belongs to the mitochondrial sirtuin family, which constitutes a highly conserved family of nicotinamide adenine dinucleotide NAD + -dependent deacetylases and ADP-ribosyltransferases that play an important regulatory role in stress resistance and metabolic homeostasis. SIRT5 , a member of the mitochondrial sirtuins, has been confirmed to exhibit deacetylase, desuccinylase, and demalonylase enzymatic activities. First, we showed that SIRT5 was expressed at the highest level in the bovine testis, followed by longissimus thoracis and subcutaneous adipose tissue, using real-time quantitative PCR and mRNA levels of SIRT5 during adipocyte differentiation, which increased before the first day and then decreased rapidly. To explore the molecular regulation of bovine SIRT5 expression, we cloned a 2-kb fragment of the 5′-regulatory region and the functional proximal minimal promoter of bovine SIRT5 . Electrophoretic mobility shift assays and luciferase reporter assays identified Kruppel-like factor 2 (KLF2), CCAAT enhancer binding protein beta (CEBPβ), peroxisome proliferator-activated receptor alpha (PPARα), myogenic differentiation 1 (MYOD), and nuclear respiratory factor 1 (NRF1) binding sites as transcriptional activators or repressors in the core promoter region of SIRT5 . In brief, our study focused on the mechanism underlying the transcriptional regulation of SIRT5 expression in bovine adipocytes.