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Int. J. Mol. Sci. 2018, 19(1), 213; https://doi.org/10.3390/ijms19010213

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

1
College of Animal Science and Technology, Northwest A&F University, Yangling 712100, Shaanxi, China
2
National Beef Cattle Improvement Center, Northwest A&F University, Yangling 712100, Shaanxi, China
*
Author to whom correspondence should be addressed.
Received: 5 December 2017 / Revised: 2 January 2018 / Accepted: 9 January 2018 / Published: 16 January 2018
(This article belongs to the Section Biochemistry, Molecular and Cellular Biology)
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Abstract

DNA methylation is a major epigenetic modification of the genome and has an essential role in muscle development. The SIX1 gene is thought to play a principal role in mediating skeletal muscle development. In the present study, we determined that bovine SIX1 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 SIX1 gene exhibited significantly higher DNA methylation levels in the AB and differentiated QCMCs groups. In addition, we found that DNA methylation of SIX1 core promoter in vitro 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 SIX1 methylation regulation during development. The results of this study provide the foundation for a better understanding of the regulation of bovine SIX1 expression via methylation and muscle developmental in beef cattle. View Full-Text
Keywords: SIX1 gene; promoter; DNA methylation; histone H4; E2F2 SIX1 gene; promoter; DNA methylation; histone H4; E2F2
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. (CC BY 4.0).
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Wei, D.; Li, A.; Zhao, C.; Wang, H.; Mei, C.; Khan, R.; Zan, L. Transcriptional Regulation by CpG Sites Methylation in the Core Promoter Region of the Bovine SIX1 Gene: Roles of Histone H4 and E2F2. Int. J. Mol. Sci. 2018, 19, 213.

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