title: Human TMEM30a promotes uptake of antitumor and bioactive choline phospholipids into mammalian cells.
abstract: Antitumor alkylphospholipids initiate apoptosis in transformed HL-60 and Jurkat cells while sparing their progenitors. 1-O-Alkyl-2-carboxymethyl-sn-glycero-3-phosphocholine (Edelfosine) like other short-chained phospholipids--inflammatory platelet-activating factor (PAF) and apoptotic oxidatively truncated phospholipids--are proposed to have intracellular sites of action, yet a conduit for these choline phospholipids into mammalian cells is undefined. Edelfosine is also accumulated by Saccharomyces cerevisiae in a process requiring the membrane protein Lem3p, and the human genome contains a Lem3p homolog TMEM30a. We show that import of choline phospholipids into S. cerevisiae ΔLem3 is partially reconstituted by human TMEM30a and by Lem3p-TMEM30a chimeras, showing the proteins are orthologous. TMEM30a-GFP chimeras expressed in mammalian cells localized in plasma membranes, as well as internal organelles, and ectopic TMEM30a expression promoted uptake of exogenous choline and ethanolamine phospholipids. Short hairpin RNA knockdown of TMEM30a reduced fluorescent choline phospholipid and [(3)H]PAF import. This knockdown also reduced mitochondrial depolarization from exogenous Edelfosine or the mitotoxic oxidatively truncated phospholipid azelaoyl phosphatidylcholine, and the knockdown reduced apoptosis in response to these two phospholipids. These results show that extracellular choline phospholipids with short sn-2 residues can have intracellular roles and sites of metabolism because they are transport substrates for a TMEM30a phospholipid import system. Variation in this mechanism could limit sensitivity to short chain choline phospholipids such as Edelfosine, PAF, and proapoptotic phospholipids.
authors.affiliation: Department of Cell Biology, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Rui
authors.initials: R
authors.lastname: Chen
authors.forename: Erin
authors.initials: E
authors.lastname: Brady
authors.forename: Thomas M
authors.initials: TM
authors.lastname: McIntyre
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D000970:Antineoplastic Agents / Q000378:metabolism*; D001693:Biological Transport, Active / Q000235:genetics / Q000276:immunology; D016466:CHO Cells; D002461:Cell Line, Transformed; D002794:Choline / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D019008:Drug Resistance, Neoplasm / Q000235:genetics / Q000276:immunology; D018922:HL-60 Cells; D056945:Hep G2 Cells; D006801:Humans; D019169:Jurkat Cells; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000502:physiology*; D026901:Membrane Transport Proteins / Q000235:genetics / Q000502:physiology; D008969:Molecular Sequence Data; D050506:Mutant Chimeric Proteins / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology; D010743:Phospholipids / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000502:physiology; D017386:Sequence Homology, Amino Acid
chemical_list: D000970:Antineoplastic Agents; C427246:CDC50 protein, S cerevisiae; C478771:Lem3 protein, S cerevisiae; D008565:Membrane Proteins; D026901:Membrane Transport Proteins; D050506:Mutant Chimeric Proteins; D010743:Phospholipids; D029701:Saccharomyces cerevisiae Proteins; C526978:TMEM30a protein, human; D002794:Choline