Free fatty acids repress the GLUT4 gene expression in cardiac muscle via novel response elements.
Hyperlipidemia (HL) impairs cardiac glucose homeostasis, but the molecular mechanisms involved are yet unclear. We examined HL-regulated GLUT4 and peroxisome proliferator-activated receptor (PPAR) gamma gene expression in human cardiac muscle. Compared with control patients, GLUT4 protein levels were 30% lower in human cardiac muscle biopsies from patients with HL and/or type 2 diabetes mellitus, whereas GLUT4 mRNA levels were unchanged. PPARgamma mRNA levels were 30-50% lower in patients with HL and/or diabetes mellitus type 2 than in controls. Reporter studies in H9C2 cardiomyotubes showed that HL in vitro, induced by high levels of arachidonic (AA) stearic, linoleic, and oleic acids (24 h, 200 mum) repressed transcription from the GLUT4 promoter; AA also repressed transcription from the PPARgamma1 and PPARgamma2 promoters. Co-expression of PPARgamma2 repressed GLUT4 promoter activity, and the addition of AA further enhanced this effect. 5'-Deletion analysis revealed three GLUT4 promoter regions that accounted for AA-mediated effects: two repression-mediating sequences at -443/-423 bp and -222/-197 bp, the deletion of either or both of which led to a partial derepression of promoter activity, and a third derepression-mediating sequence at -612/-587 bp that was required for sustaining this derepression effect. Electromobility shift assay further shows that AA enhanced binding to two of the three regions of cardiac nuclear protein(s), the nature of which is still unknown. We propose that HL, exhibited as a high free fatty acid level, modulates GLUT4 gene expression in cardiac muscle via a complex mechanism that includes: (a) binding of AA mediator proteins to three newly identified response elements on the GLUT4 promoter gene and (b) repression of GLUT4 and the PPARgamma genes by AA.
Armoni
Michal
M
Institute of Endocrinology, Diabetes and Metabolism, Rambam Medical Center and B. Rappaport Faculty of Medicine, Technion-Israel Institute of Technology, Haifa 31096, Israel.
Harel
Chava
C
Bar-Yoseph
Fabiana
F
Milo
Simcha
S
Karnieli
Eddy
E
D000368:Aged; D000818:Animals; D016718:Arachidonic Acid / Q000378:metabolism / Q000494:pharmacology; D001706:Biopsy; D015153:Blotting, Western; D016466:CHO Cells; D002467:Cell Nucleus / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D006224:Cricetinae; D004247:DNA / Q000737:chemistry; D017931:DNA Primers / Q000737:chemistry; D003847:Deoxyglucose / Q000378:metabolism / Q000493:pharmacokinetics; D003924:Diabetes Mellitus, Type 2 / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D005227:Fatty Acids / Q000737:chemistry* / Q000378:metabolism; D005260:Female; D017353:Gene Deletion; D005786:Gene Expression Regulation; D017930:Genes, Reporter; D051275:Glucose Transporter Type 4 / Q000096:biosynthesis* / Q000235:genetics*; D006367:HeLa Cells; D006801:Humans; D019787:Linoleic Acid / Q000378:metabolism; D008055:Lipids / Q000737:chemistry; D008297:Male; D008875:Middle Aged; D008954:Models, Biological; D009132:Muscles / Q000166:cytology; D009206:Myocardium / Q000378:metabolism*; D019301:Oleic Acid / Q000378:metabolism; D047495:PPAR gamma / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D020218:Response Elements; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013229:Stearic Acids / Q000378:metabolism; D013997:Time Factors; D014158:Transcription, Genetic
D017931:DNA Primers; D005227:Fatty Acids; D051275:Glucose Transporter Type 4; D008055:Lipids; D047495:PPAR gamma; D011506:Proteins; D012333:RNA, Messenger; D013229:Stearic Acids; D016718:Arachidonic Acid; D019301:Oleic Acid; C031183:stearic acid; D004247:DNA; D003847:Deoxyglucose; D019787:Linoleic Acid