title: Diacylglycerol acyltransferase-2 (DGAT2) and monoacylglycerol acyltransferase-2 (MGAT2) interact to promote triacylglycerol synthesis.
abstract: Acyl CoA:1,2-diacylglycerol acyltransferase (DGAT)-2 is an integral membrane protein that catalyzes triacylglycerol (TG) synthesis using diacylglycerol and fatty acyl CoA as substrates. DGAT2 resides in the endoplasmic reticulum (ER), but when cells are incubated with fatty acids, DGAT2 interacts with lipid droplets presumably to catalyze localized TG synthesis for lipid droplet expansion. Previous studies have shown that DGAT2 interacts with proteins that synthesize its fatty acyl CoA substrates. In this study, we provide additional evidence that DGAT2 is present in a protein complex. Using a chemical cross-linker, disuccinimidyl suberate (DSS), we demonstrated that DGAT2 formed a dimer and was also part of a protein complex of ∼ 650 kDa, both in membranes and on lipid droplets. Using co-immunoprecipitation experiments and an in situ proximity ligation assay, we found that DGAT2 interacted with monoacylglycerol acyltransferase (MGAT)-2, an enzyme that catalyzes the synthesis of diacylglycerol. Deletion mutagenesis showed that the interaction with MGAT2 was dependent on the two transmembrane domains of DGAT2. No significant interaction of DGAT2 with lipin1, another enzyme that synthesizes diacylglycerol, could be detected. When co-expressed in cells, DGAT2 and MGAT2 co-localized in the ER and on lipid droplets. Co-expression also resulted in increased TG storage compared with expression of DGAT2 or MGAT2 alone. Incubating McArdle rat hepatoma RH7777 cells with 2-monoacylglycerol caused DGAT2 to translocate to lipid droplets. This also led to the formation of large cytosolic lipid droplets, characteristic of DGAT2, but not DGAT1, and indicated that DGAT2 can utilize monoacylglycerol-derived diacylglycerol. These findings suggest that the interaction of DGAT2 and MGAT2 serves to channel lipid substrates efficiently for TG biosynthesis.
authors.affiliation: From the Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada and.
authors.forename: Youzhi
authors.initials: Y
authors.lastname: Jin
authors.affiliation: From the Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada and.
authors.forename: Pamela J
authors.initials: PJ
authors.lastname: McFie
authors.affiliation: Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5B4, Canada.
authors.forename: Shanna L
authors.initials: SL
authors.lastname: Banman
authors.affiliation: From the Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada and.
authors.forename: Curtis
authors.initials: C
authors.lastname: Brandt
authors.affiliation: From the Department of Biochemistry, University of Saskatchewan, Saskatoon, Saskatchewan S7N 5E5, Canada and scot.stone@usask.ca.
authors.forename: Scot J
authors.initials: SJ
authors.lastname: Stone
mesh_terms: D000217:Acyltransferases / Q000235:genetics* / Q000378:metabolism; D000818:Animals; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D003432:Cross-Linking Reagents / Q000737:chemistry; D051048:Diacylglycerol O-Acyltransferase / Q000235:genetics* / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000737:chemistry / Q000378:metabolism; D005786:Gene Expression Regulation; D057809:HEK293 Cells; D022781:Hepatocytes / Q000166:cytology / Q000201:enzymology*; D006801:Humans; D066292:Lipid Droplets / Q000737:chemistry / Q000378:metabolism; D050178:Monoglycerides / Q000378:metabolism; D011485:Protein Binding; D055503:Protein Multimerization; D051381:Rats; D015398:Signal Transduction; D013388:Succinimides / Q000737:chemistry; D014280:Triglycerides / Q000096:biosynthesis*
chemical_list: D003432:Cross-Linking Reagents; D050178:Monoglycerides; D013388:Succinimides; D014280:Triglycerides; D000217:Acyltransferases; C522579:Dgat2 protein, rat; D051048:Diacylglycerol O-Acyltransferase; C021802:2-acylglycerol O-acyltransferase; C019358:disuccinimidyl suberate
keywords: Diacylglycerol Acyltransferase; Lipid; Lipid Droplet; Monoacylglycerol Acyltransferase; Protein Cross-linking; Triacylglycerol; diacylglycerol