Characterization of the mouse and human Monoacylglycerol O-Acyltransferase 1 (Mogat1) promoter in human kidney proximal tubule and rat liver cells

Shireesha Sankella, Abhimanyu Garg, Anil K. Agarwal

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5 Citations (Scopus)

Abstract

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 reportergene activity. Chromatin immunoprecipitation (ChIP) assay using PPARα antibody further confirmedthe 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.

Original languageEnglish (US)
Article numbere0162504
JournalPLoS One
Volume11
Issue number9
DOIs
StatePublished - Sep 1 2016

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Monoglycerides
Acyltransferases
Proximal Kidney Tubule
acyltransferases
proximal tubules
monoacylglycerols
Liver
hepatocytes
Rats
promoter regions
kidneys
Peroxisome Proliferator-Activated Receptors
rats
mice
Peroxisome Proliferators
Assays
Response Elements
Tissue
response elements
peroxisomes

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)

Cite this

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title = "Characterization of the mouse and human Monoacylglycerol O-Acyltransferase 1 (Mogat1) promoter in human kidney proximal tubule and rat liver cells",
abstract = "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 reportergene activity. Chromatin immunoprecipitation (ChIP) assay using PPARα antibody further confirmedthe 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.",
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