title: Identification of a novel lysophospholipid acyltransferase in Saccharomyces cerevisiae.
abstract: The incorporation of unsaturated acyl chains into phospholipids during de novo synthesis is primarily mediated by the 1-acyl-sn-glycerol-3-phosphate acyltransferase reaction. In Saccharomyces cerevisiae, Slc1 has been shown to mediate this reaction, but distinct activity remains after its removal from the genome. To identify the enzyme that mediates the remaining activity, we performed synthetic genetic array analysis using a slc1Delta strain. One of the genes identified by the screen, LPT1, was found to encode for an acyltransferase that uses a variety of lysophospholipid species, including 1-acyl-sn-glycerol-3-phosphate. Deletion of LPT1 had a minimal effect on 1-acyl-sn-glycerol-3-phosphate acyltransferase activity, but overexpression increased activity 7-fold. Deletion of LPT1 abrogated the esterification of other lysophospholipids, and overexpression increased lysophosphatidylcholine acyltransferase activity 7-fold. The majority of this activity co-purified with microsomes. To test the putative role for this enzyme in selectively incorporating unsaturated acyl chains into phospholipids in vitro, substrate concentration series experiments were performed with the four acyl-CoA species commonly found in yeast. Although the saturated palmitoyl-CoA and stearoyl-CoA showed a lower apparent Km, the monounsaturated palmitoleoyl-CoA and oleoyl-CoA showed a higher apparent Vmax. Arachidonyl-CoA, although not abundant in yeast, also had a high apparent Vmax. Pulse-labeling of lpt1Delta strains showed a 30% reduction in [3H]oleate incorporation into phosphatidylcholine only. Therefore, Lpt1p, a member of the membrane-bound o-acyltransferase gene family, seems to work in conjunction with Slc1 to mediate the incorporation of unsaturated acyl chains into the sn-2 position of phospholipids.
authors.affiliation: Department of Bioscience and Biotechnology, Drexel University, Philadelphia, Pennsylvania 19104, USA.
authors.forename: Shilpa
authors.initials: S
authors.lastname: Jain
authors.forename: NaTaza
authors.initials: N
authors.lastname: Stanford
authors.forename: Neha
authors.initials: N
authors.lastname: Bhagwat
authors.forename: Brian
authors.initials: B
authors.lastname: Seiler
authors.forename: Michael
authors.initials: M
authors.lastname: Costanzo
authors.forename: Charles
authors.initials: C
authors.lastname: Boone
authors.forename: Peter
authors.initials: P
authors.lastname: Oelkers
mesh_terms: D008243:1-Acylglycerophosphocholine O-Acyltransferase / Q000235:genetics / Q000378:metabolism*; D000217:Acyltransferases / Q000235:genetics / Q000378:metabolism*; D002462:Cell Membrane / Q000201:enzymology* / Q000235:genetics; D003067:Coenzymes / Q000378:metabolism; D004398:Dyneins; D005231:Fatty Acids, Unsaturated / Q000235:genetics / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology; D015966:Gene Expression Regulation, Fungal / Q000502:physiology; D007700:Kinetics; D008244:Lysophosphatidylcholines / Q000096:biosynthesis* / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D012441:Saccharomyces cerevisiae / Q000201:enzymology* / Q000235:genetics; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: D003067:Coenzymes; D005231:Fatty Acids, Unsaturated; D008244:Lysophosphatidylcholines; D029701:Saccharomyces cerevisiae Proteins; D000217:Acyltransferases; D008243:1-Acylglycerophosphocholine O-Acyltransferase; D004398:Dyneins; C083745:SLC1 protein, S cerevisiae