title: ORMDL/serine palmitoyltransferase stoichiometry determines effects of ORMDL3 expression on sphingolipid biosynthesis.
abstract: The ORM1 (Saccharomyces cerevisiae)-like proteins (ORMDLs) and their yeast orthologs, the Orms, are negative homeostatic regulators of the initiating enzyme in sphingolipid biosynthesis, serine palmitoyltransferase (SPT). Genome-wide association studies have established a strong correlation between elevated expression of the endoplasmic reticulum protein ORMDL3 and risk for childhood asthma. Here we test the notion that elevated levels of ORMDL3 decrease sphingolipid biosynthesis. This was tested in cultured human bronchial epithelial cells (HBECs) (an immortalized, but untransformed, airway epithelial cell line) and in HeLa cells (a cervical adenocarcinoma cell line). Surprisingly, elevated ORMDL3 expression did not suppress de novo biosynthesis of sphingolipids. We determined that ORMDL is expressed in functional excess relative to SPT at normal levels of expression. ORMDLs and SPT form stable complexes that are not increased by elevated ORMDL3 expression. Although sphingolipid biosynthesis was not decreased by elevated ORMDL3 expression, the steady state mass levels of all major sphingolipids were marginally decreased by low level ORMDL3 over-expression in HBECs. These data indicate that the contribution of ORMDL3 to asthma risk may involve changes in sphingolipid metabolism, but that the connection is complex.
authors.affiliation: James Graham Brown Cancer Center University of Louisville School of Medicine, Louisville, KY 40202.
authors.forename: Deanna
authors.initials: D
authors.lastname: Siow
authors.affiliation: Division of Cardiovascular Medicine, Gill Heart Institute, University of Kentucky Lexington and Department of Veterans Affairs Medical Center, Lexington, KY 40536.
authors.forename: Manjula
authors.initials: M
authors.lastname: Sunkara
authors.affiliation: Department of Biochemistry, Uniformed Services University of the Health Sciences, Bethesda, MD 20184.
authors.forename: Teresa M
authors.initials: TM
authors.lastname: Dunn
authors.affiliation: Division of Cardiovascular Medicine, Gill Heart Institute, University of Kentucky Lexington and Department of Veterans Affairs Medical Center, Lexington, KY 40536.
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Morris
authors.affiliation: James Graham Brown Cancer Center University of Louisville School of Medicine, Louisville, KY 40202 Departments of Medicine, Biochemistry, and Molecular Biology, Pharmacology, and Toxicology, University of Louisville School of Medicine, Louisville, KY 40202.
authors.forename: Binks
authors.initials: B
authors.lastname: Wattenberg
mesh_terms: D000818:Animals; D001249:Asthma / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D001980:Bronchi / Q000166:cytology; D004847:Epithelial Cells / Q000378:metabolism; D015870:Gene Expression; D020868:Gene Silencing; D006367:HeLa Cells; D006801:Humans; D008565:Membrane Proteins / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism*; D051379:Mice; D010641:Phenotype; D034741:RNA, Small Interfering / Q000235:genetics; D051102:Serine C-Palmitoyltransferase / Q000378:metabolism*; D013107:Sphingolipids / Q000096:biosynthesis* / Q000378:metabolism
chemical_list: D008565:Membrane Proteins; C464399:ORMDL3 protein, human; D034741:RNA, Small Interfering; D013107:Sphingolipids; D051102:Serine C-Palmitoyltransferase
keywords: ORM1 (Saccharomyces cerevisiae)-like protein; Orm; asthma; ceramides; endoplasmic reticulum; enzymology/enzyme regulation; lung; sphingosine phosphate