Characterization of the Mouse and Human Monoacylglycerol O-Acyltransferase 1 (Mogat1) Promoter in Human Kidney Proximal Tubule and Rat Liver Cells.
Monoacylglycerol acyltransferase 1 (Mogat1) catalyzes the conversion of monoacylglycerols (MAG) to diacylglycerols (DAG), the precursor of several physiologically important lipids such as phosphatidylcholine, phosphatidylethanolamine and triacylglycerol (TAG). Expression of Mogat1 is tissue restricted and it is highly expressed in the kidney, stomach and adipose tissue but minimally in the normal adult liver. To understand the transcriptional regulation of Mogat1, we characterized the mouse and human Mogat1 promoters in human kidney proximal tubule-2 (HK-2) cells. In-silico analysis revealed several peroxisome proliferator response element (PPRE) binding sites in the promoters of both human and mouse Mogat1. These sites responded to all three peroxisome proliferator activated receptor (PPAR) isoforms such that their respective agonist or antagonist activated or inhibited the expression of Mogat1. PPRE site mutagenesis revealed that sites located at -592 and -2518 are very effective in decreasing luciferase reporter gene activity. Chromatin immunoprecipitation (ChIP) assay using PPARα antibody further confirmed the occupancy of these sites by PPARα. While these assays revealed the core promoter elements necessary for Mogat1 expression, there are additional elements required to regulate its tissue specific expression. Chromosome conformation capture (3C) assay revealed additional cis-elements located ~10-15 kb upstream which interact with the core promoter. These chromosomal regions are responsive to both PPARα agonist and antagonist.
Division of Nutrition and Metabolic Diseases, Department of Internal Medicine and Center for Human Nutrition, UT Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390, United States of America.
Shireesha
S
Sankella
Division of Nutrition and Metabolic Diseases, Department of Internal Medicine and Center for Human Nutrition, UT Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390, United States of America.
Abhimanyu
A
Garg
Division of Nutrition and Metabolic Diseases, Department of Internal Medicine and Center for Human Nutrition, UT Southwestern Medical Center, 5323 Harry Hines Blvd., Dallas, Texas 75390, United States of America.
Anil K
AK
Agarwal
D000217:Acyltransferases / Q000235:genetics*; D000813:Anilides / Q000494:pharmacology; D000818:Animals; D015152:Blotting, Northern; D018938:Caco-2 Cells; D002460:Cell Line; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D019295:Computational Biology; D019073:HT29 Cells; D006801:Humans; D007687:Kidney Tubules, Proximal / Q000187:drug effects / Q000378:metabolism; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D010080:Oxazoles / Q000494:pharmacology; D047493:PPAR alpha / Q000819:agonists / Q000037:antagonists & inhibitors / Q000378:metabolism; D011401:Promoter Regions, Genetic / Q000235:genetics*; D011743:Pyrimidines / Q000494:pharmacology; D051381:Rats; D060888:Real-Time Polymerase Chain Reaction; D000077154:Rosiglitazone; D013450:Sulfones / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D045162:Thiazolidinediones / Q000494:pharmacology; D013876:Thiophenes / Q000494:pharmacology; D014443:Tyrosine / Q000031:analogs & derivatives / Q000494:pharmacology; D010636:Phenols; D013438:Sulfhydryl Compounds
D000217:Acyltransferases; D000813:Anilides; D010080:Oxazoles; D047493:PPAR alpha; D011743:Pyrimidines; D000077154:Rosiglitazone; D013450:Sulfones; D013844:Thiazoles; D045162:Thiazolidinediones; D013876:Thiophenes; D014443:Tyrosine; C457499:2-chloro-5-nitrobenzanilide; C529769:GSK0660; C449302:GW 6471; C479979:(4-(((2-(3-fluoro-4-(trifluoromethyl)phenyl)-4-methyl-1,3-thiazol-5-yl)methyl)sulfanyl)-2-methylphenoxy)acetic acid; D010636:Phenols; D013438:Sulfhydryl Compounds; C006253:pirinixic acid; C021802:2-acylglycerol O-acyltransferase