Abstract
The SIX1 homeobox gene belongs to the six homeodomain family and is widely thought to play a principal role in mediating of skeletal muscle development. In the present study, we determined that the bovine SIX1 gene was highly expressed in the longissimus thoracis and physiologically immature individuals. DNA sequencing of 428 individual Qinchuan cattle identified nine single nucleotide polymorphisms (SNPs) in the promoter region of the SIX1 gene. Using a series of 5′ deletion promoter plasmid luciferase reporter assays and 5′-rapid amplification of cDNA end analysis (RACE), two of these SNPs were found to be located in the proximal minimal promoter region −216/−28 relative to the transcriptional start site (TSS). Correlation analysis showed the combined haplotypes H1-H2 (-GG-GA-) was significantly greater in the body measurement traits (BMTs) than the others, which was consistent with the results showing that the transcriptional activity of Hap2 was higher than the others in Qinchuan cattle myoblast cells. Furthermore, the electrophoretic mobility shift assays (EMSA) and chromatin immunoprecipitation assay (ChIP) demonstrated that NRF1 and ZSCAN10 binding occurred in the promoter region of diplotypes H1-H2 to regulate SIX1 transcriptional activity. This information may be useful for molecular marker-assisted selection (MAS) in cattle breeding.
Highlights
Body measurement traits (BMTs) are relevant indicators in cattle selection and breeding
Our work supports that sine oculis homeobox homologue 1 (SIX1) is one of the genes that controls myogenesis and may influence body measurement traits (BMTs)
The tissue distribution of SIX1 mRNA showed the highest expression in the longissimus thoracis (Fig. 1a), which is consistent with previous findings observed in other species, such as in human[22], porcine[23] and duck[24]
Summary
Body measurement traits (BMTs) are relevant indicators in cattle selection and breeding. It is necessary to select important functional genes to solve these problems and increase the BMTs of beef cattle through marker-assisted selection (MAS), which is recognized as a powerful and efficient strategy compared to traditional breeding methods[1]. Quantitative traits such as BMTs are controlled by many genes with minor effects[2]. Despite its clear role in regulating the formation of muscle and other tissues, the mechanism by which the bovine SIX1 gene is associated with BMTs in cattle has not been reported to date. The objectives of this study were to identify the genetic polymorphisms at the 5′UTR of the bovine SIX1 gene and confirm potential cis-acting elements that are associated with single nucleotide polymorphisms (SNPs) and BMTs in Qinchuan cattle
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