title: Characterization of the promoter region of the bovine long-chain acyl-CoA synthetase 1 gene: Roles of E2F1, Sp1, KLF15, and E2F4.
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 integrate and frame a key functional role for ACSL1 in mediating the lipid composition of beef.
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: Zhi-Dong
authors.initials: ZD
authors.lastname: Zhao
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.|National Beef Cattle Improvement Center, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: Lin-Sen
authors.initials: LS
authors.lastname: Zan
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: An-Ning
authors.initials: AN
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.|National Beef Cattle Improvement Center, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: Gong
authors.initials: G
authors.lastname: Cheng
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: Shi-Jun
authors.initials: SJ
authors.lastname: Li
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: Ya-Ran
authors.initials: YR
authors.lastname: Zhang
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: Xiao-Yu
authors.initials: XY
authors.lastname: Wang
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling 712100 Shaanxi, People's Republic of China.
authors.forename: Ying-Ying
authors.initials: YY
authors.lastname: Zhang
mesh_terms: D000273:Adipose Tissue / Q000378:metabolism; D000818:Animals; D001483:Base Sequence; D002417:Cattle; D003066:Coenzyme A Ligases / Q000235:genetics*; D018899:CpG Islands / Q000235:genetics; D004252:DNA Mutational Analysis / Q000379:methods; D050687:E2F1 Transcription Factor / Q000235:genetics*; D050704:E2F4 Transcription Factor / Q000235:genetics*; D005231:Fatty Acids, Unsaturated / Q000235:genetics; D051741:Kruppel-Like Transcription Factors / Q000235:genetics*; D008055:Lipids / Q000235:genetics; D018482:Muscle, Skeletal / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011485:Protein Binding / Q000235:genetics; D000069466:Red Meat; D017384:Sequence Deletion / Q000235:genetics; D016329:Sp1 Transcription Factor / Q000235:genetics*; D024363:Transcription Initiation Site / Q000502:physiology; D014158:Transcription, Genetic / Q000235:genetics
chemical_list: D050687:E2F1 Transcription Factor; D050704:E2F4 Transcription Factor; D005231:Fatty Acids, Unsaturated; D051741:Kruppel-Like Transcription Factors; D008055:Lipids; D016329:Sp1 Transcription Factor; D003066:Coenzyme A Ligases