Cloning and expression of mouse liver phosphatidylserine synthase-1 cDNA. Overexpression in rat hepatoma cells inhibits the CDP-ethanolamine pathway for phosphatidylethanolamine biosynthesis.
In eukaryotic cells, phosphatidylserine (PtdSer) is synthesized by two distinct synthases on the endoplasmic reticulum by a base-exchange reaction in which the polar head-group of an existing phospholipid is replaced with serine. We report the cloning and expression of a cDNA for mouse liver PtdSer synthase-1. The deduced protein sequence is >90% identical to that of PtdSer synthase-1 from Chinese hamster ovary cells and a sequence from a human myeloblast cell line. PtdSer synthase-1 cDNA was stably expressed in M.9.1.1 cells which are mutant Chinese hamster ovary cells defective in PtdSer synthase-1 activity, are ethanolamine auxotrophs, and have a reduced content of PtdSer and phosphatidylethanolamine (PtdEtn). The growth defect of M.9.1.1 cells was eliminated, and a normal phospholipid composition was restored in the absence of exogenous ethanolamine, implying that the cloned cDNA encoded PtdSer synthase. Mouse liver PtdSer synthase-1 was also expressed in McArdle 7777 rat hepatoma cells. In addition to a 3-fold higher in vitro serine-exchange activity, these cells also exhibited enhanced choline- and ethanolamine-exchange activities and incorporated more [3H]serine into PtdSer than did control cells. However, the levels of PtdSer and PtdEtn in cells overexpressing PtdSer synthase-1 activity were not increased. Excess PtdSer produced by the transfected cells was rapidly decarboxylated to PtdEtn and the degradation of PtdSer, and/or PtdEtn derived from PtdSer, was increased. Moreover, the CDP-ethanolamine pathway for PtdEtn biosynthesis was inhibited. These data suggest that (i) cellular levels of PtdSer and PtdEtn are tightly controlled, and (ii) the metabolism of PtdSer and PtdEtn is coordinately regulated to maintain phospholipid homeostasis.
Stone
S J
SJ
Lipid and Lipoprotein Research Group, University of Alberta, Edmonton, AB T6G 2S2, Canada.
Cui
Z
Z
Vance
J E
JE
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D016466:CHO Cells; D006528:Carcinoma, Hepatocellular; D003001:Cloning, Molecular; D006224:Cricetinae; D003565:Cytidine Diphosphate / Q000031:analogs & derivatives* / Q000378:metabolism; D018076:DNA, Complementary / Q000737:chemistry / Q000302:isolation & purification; D004983:Ethanolamines / Q000378:metabolism*; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D008099:Liver / Q000201:enzymology*; D008565:Membrane Proteins / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D019884:Nitrogenous Group Transferases / Q000235:genetics*; D010714:Phosphatidylethanolamines / Q000096:biosynthesis*; D010718:Phosphatidylserines / Q000096:biosynthesis*; D051381:Rats; D012694:Serine / Q000378:metabolism; D014407:Tumor Cells, Cultured
D018076:DNA, Complementary; D004983:Ethanolamines; D008565:Membrane Proteins; D010714:Phosphatidylethanolamines; D010718:Phosphatidylserines; C006933:CDP ethanolamine; D012694:Serine; D003565:Cytidine Diphosphate; D019884:Nitrogenous Group Transferases; C111493:Ptdss2 protein, mouse; C022370:phospholipid serine base exchange enzyme