Characterization of the promoter region of the bovine long-chain acyl-CoA synthetase 1 gene: Roles of E2F1, Sp1, KLF15, and E2F4

Scientific Reports
Zhi-Dong ZhaoYing-Ying Zhang

Abstract

The nutritional value and eating qualities of beef are enhanced when the unsaturated fatty acid content of fat is increased. Long-chain acyl-CoA synthetase 1 (ACSL1) plays key roles in fatty acid transport and degradation, as well as lipid synthesis. It has been identified as a plausible functional and positional candidate gene for manipulations of fatty acid composition in bovine skeletal muscle. In the present study, we determined that bovine ACSL1 was highly expressed in subcutaneous adipose tissue and longissimus thoracis. To elucidate the molecular mechanisms involved in bovine ACSL1 regulation, we cloned and characterized the promoter region of ACSL1. Applying 5'-rapid amplification of cDNA end analysis (RACE), we identified multiple transcriptional start sites (TSSs) in its promoter region. Using a series of 5' deletion promoter plasmids in luciferase reporter assays, we found that the proximal minimal promoter of ACSL1 was located within the region -325/-141 relative to the TSS and it was also located in the predicted CpG island. Mutational analysis and electrophoretic mobility shift assays demonstrated that E2F1, Sp1, KLF15 and E2F4 binding to the promoter region drives ACSL1 transcription. Together these interactions ...Continue Reading

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Citations

Sep 8, 2019·Animals : an Open Access Journal From MDPI·Zhidong ZhaoJiang Hu
Sep 7, 2019·International Journal of Molecular Sciences·Rajwali KhanZan Linsen
Aug 11, 2020·Molecular Reproduction and Development·Mohamed AshryGeorge W Smith
Jan 13, 2019·International Journal of Molecular Sciences·Girmay ShishayChenghui Liu
Oct 26, 2020·Archives of Biochemistry and Biophysics·Zhidong ZhaoJiang Hu

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Methods Mentioned

BETA
PCR
transfection
electrophoretic mobility shift
histone acetylation
electrophoresis

Software Mentioned

Matlnspector
MatInspector
MethPrimer

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