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TNF-? induces acyl-CoA synthetase 3 to promote lipid droplet formation in human endothelial cells.

ABSTRACT: Chronic inflammation contributes to cardiovascular disease. Increased levels of the inflammatory cytokine, TNF-?, are often present in conditions associated with cardiovascular disease risk, and TNF-? induces a number of pro-atherogenic effects in macrovascular endothelial cells, including expression of adhesion molecules and chemokines, and lipoprotein uptake and transcytosis to the subendothelial tissue. However, little is known about the roles of acyl-CoA synthetases (ACSLs), enzymes that esterify free fatty acids into their acyl-CoA derivatives, or about the effects of TNF-? on ACSLs in endothelial cells. Therefore, we investigated the effects of TNF-? on ACSLs and downstream lipids in cultured human coronary artery endothelial cells and human umbilical vein endothelial cells. We demonstrated that TNF-? induces ACSL1, ACSL3, and ACSL5, but not ACSL4, in both cell types. TNF-? also increased oleoyl-CoA levels, consistent with the increased ACSL3 expression. RNA-sequencing demonstrated that knockdown of ACSL3 had no marked effects on the TNF-? transcriptome. Instead, ACSL3 was required for TNF-?-induced lipid droplet formation in cells exposed to oleic acid. These results demonstrate that increased acyl-CoA synthesis as a result of ACSL3 induction is part of the TNF-? response in human macrovascular endothelial cells.

SUBMITTER: Jung HS

PROVIDER: S-EPMC6939593 | biostudies-literature | 2020 Jan

REPOSITORIES: biostudies-literature

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TNF-α induces acyl-CoA synthetase 3 to promote lipid droplet formation in human endothelial cells.

Jung Hye Seung HS Shimizu-Albergine Masami M Shen Xia X Kramer Farah F Shao Dan D Vivekanandan-Giri Anuradha A Pennathur Subramaniam S Tian Rong R Kanter Jenny E JE Bornfeldt Karin E KE

Journal of lipid research 20191113 1

Chronic inflammation contributes to cardiovascular disease. Increased levels of the inflammatory cytokine, TNF-α, are often present in conditions associated with cardiovascular disease risk, and TNF-α induces a number of pro-atherogenic effects in macrovascular endothelial cells, including expression of adhesion molecules and chemokines, and lipoprotein uptake and transcytosis to the subendothelial tissue. However, little is known about the roles of acyl-CoA synthetases (ACSLs), enzymes that est ...[more]

PMID: 31722970

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Project description:Long chain acyl-CoA synthetase 1 (ACSL1) contributes 50 to 90% of total ACSL activity in liver, adipose tissue, and heart and appears to direct the use of long chain fatty acids for energy. Although the functional importance of ACSL1 is becoming clear, little is understood about its post-translational regulation. In order to investigate the post-translational modifications of ACSL1 under different physiological conditions, we overexpressed ACSL1 in hepatocytes, brown adipocytes, and 3T3-L1 differentiated adipocytes, treated these cells with different hormones, and analyzed the resulting phosphorylated and acetylated amino acids by mass spectrometry. We then compared these results to the post-translational modifications observed in vivo in liver and brown adipose tissue after mice were fasted or exposed to a cold environment. We identified universal N-terminal acetylation, 15 acetylated lysines, and 25 phosphorylation sites on ACSL1. Several unique acetylation and phosphorylation sites occurred under conditions in which fatty acid β-oxidation is normally enhanced. Thirteen of the acetylated lysines had not previously been identified, and none of the phosphorylation sites had been previously identified. Site-directed mutagenesis was used to introduce mutations at three potential acetylation and phosphorylation sites believed to be important for ACSL1 function. At the ATP/AMP binding site and at a highly conserved site near the C terminus, modifications of Ser278 or Lys676, respectively, totally inhibited ACSL1 activity. In contrast, mutations of Lys285 that mimicked acetylation (Lys285Ala and Lys285Gln) reduced ACSL activity, whereas full activity was retained by Lys285Arg, suggesting that acetylation of Lys285 would be likely to decrease ACSL1 activity. These results indicate that ACSL1 is highly modified post-translationally. Several of these modifications would be expected to alter enzymatic function, but others may affect protein stability or protein-protein interactions.

Dataset Information

TNF-? induces acyl-CoA synthetase 3 to promote lipid droplet formation in human endothelial cells.

Publications

TNF-α induces acyl-CoA synthetase 3 to promote lipid droplet formation in human endothelial cells.

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