Functional analysis of long-chain acyl-CoA synthetase 1 in 3T3-L1 adipocytes.
ACSL1 (acyl-CoA synthetase 1), the major acyl-CoA synthetase of adipocytes, has been proposed to function in adipocytes as mediating free fatty acid influx, esterification, and storage as triglyceride. To test this hypothesis, ACSL1 was stably silenced (knockdown (kd)) in 3T3-L1 cells, differentiated into adipocytes, and evaluated for changes in lipid metabolism. Surprisingly, ACSL1-silenced adipocytes exhibited no significant changes in basal or insulin-stimulated long-chain fatty acid uptake, lipid droplet size, or tri-, di-, or monoacylglycerol levels when compared with a control adipocyte line. However, ACSL1 kd adipocytes displayed a 7-fold increase in basal and a approximately 15% increase in forskolin-stimulated fatty acid efflux without any change in glycerol release, indicating a role for the protein in fatty acid reesterification following lipolysis. Consistent with this proposition, ACSL1 kd cells exhibited a decrease in activation and phosphorylation of AMP-activated protein kinase and its primary substrate acetyl-CoA carboxylase. Moreover, ACSL1 kd adipocytes displayed an increase in phosphorylated protein kinase C and phosphorylated JNK, attenuated insulin signaling, and a decrease in insulin-stimulated glucose uptake. These findings identify a primary role of ACSL1 in adipocytes not in control of lipid influx, as previously considered, but in lipid efflux and fatty acid-induced insulin resistance.
Department of Biochemistry, Molecular Biology and Biophysics, The University of Minnesota, Minneapolis, Minnesota 55455, USA.
Sandra
S
Lobo
Brian M
BM
Wiczer
David A
DA
Bernlohr
D041721:3T3-L1 Cells; D000214:Acyl Coenzyme A / Q000378:metabolism*; D000263:Adenylate Kinase / Q000378:metabolism; D017667:Adipocytes / Q000166:cytology / Q000187:drug effects / Q000201:enzymology*; D000818:Animals; D003066:Coenzyme A Ligases / Q000235:genetics / Q000378:metabolism*; D005576:Colforsin / Q000494:pharmacology; D004951:Esterification / Q000502:physiology; D005227:Fatty Acids / Q000493:pharmacokinetics; D020868:Gene Silencing; D005947:Glucose / Q000493:pharmacokinetics; D007328:Insulin / Q000378:metabolism; D007333:Insulin Resistance / Q000502:physiology; D050356:Lipid Metabolism / Q000502:physiology*; D008066:Lipolysis / Q000502:physiology*; D051379:Mice; D015398:Signal Transduction / Q000502:physiology; D014316:Tritium
D000214:Acyl Coenzyme A; D005227:Fatty Acids; D007328:Insulin; D014316:Tritium; D005576:Colforsin; D000263:Adenylate Kinase; C544400:ACSL1 protein, mouse; D003066:Coenzyme A Ligases; D005947:Glucose