Transcriptional Regulation by CpG Sites Methylation in the Core Promoter Region of the Bovine SIX1 Gene: Roles of Histone H4 and E2F2

• Published in: International journal of molecular sciences Vol. 19; no. 1; p. 213
• Main Authors: Wei, Dawei, Li, Anning, Zhao, Chunping, Wang, Hongbao, Mei, Chugang, Khan, Rajwali, Zan, Linsen
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 16.01.2018; MDPI
• Subjects: Bisulfite; Bovidae; Cattle; Chromatin; CpG islands; Deoxyribonucleic acid; DNA; DNA methylation; DNA sequencing; E2F2; Electrophoretic mobility; Fetuses; Gene regulation; Genomes; Histone H4; Immunoprecipitation; Muscles; Polymerase chain reaction; promoter; siRNA; SIX gene family; SIX1 gene; Skeletal muscle; Transcription; DNA methylation; E2F2; promoter; SIX1 gene; histone H4
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms19010213

DNA methylation is a major epigenetic modification of the genome and has an essential role in muscle development. The gene is thought to play a principal role in mediating skeletal muscle development. In the present study, we determined that bovine expression levels were significantly higher in the fetal bovine group (FB) and in undifferentiated Qinchuan cattle muscle cells (QCMCs) than in the adult bovine group (AB) and in differentiated QCMCs. Moreover, a bisulfite sequencing polymerase chain reaction (BSP) analysis of DNA methylation levels showed that three CpG sites in the core promoter region (-216/-28) of the bovine gene exhibited significantly higher DNA methylation levels in the AB and differentiated QCMCs groups. In addition, we found that DNA methylation of core promoter obviously influences the promoter activities. An electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) assay, in combination with site-directed mutation and siRNA interference, demonstrated that histone H4 and E2F2 bind to the -216/-28 region and play important roles in methylation regulation during development. The results of this study provide the foundation for a better understanding of the regulation of bovine expression via methylation and muscle developmental in beef cattle.