title: The yeast and mammalian isoforms of phosphatidylinositol transfer protein can all restore phospholipase C-mediated inositol lipid signaling in cytosol-depleted RBL-2H3 and HL-60 cells.
abstract: The mammalian phosphatidylinositol transfer proteins (PITP) and the yeast Saccharomyces cerevisiae PITP (SEC14p) that show no sequence homology both catalyze exchange of phosphatidylinositol (PI) between membranes compartments in vitro. In HL-60 cells where the cytosolic proteins are depleted by permeabilization, exogenously added PITPalpha is required to restore G protein-mediated phospholipase Cbeta (PLCbeta) signaling. Recently, a second mammalian PITPbeta form has been described that shows 77% identity to rat PITPalpha. We have examined the ability of the two mammalian PITPs and SEC14p to restore PLC-mediated signaling in cytosol-depleted HL-60 and RBL-2H3 cells. Both PITPalpha and PITPbeta isoforms as well as SEC14p restore G protein-mediated PLCbeta signaling with a similar potency. In RBL-2H3 cells, crosslinking of the IgE receptor by antigen stimulates inositol lipid hydrolysis by tyrosine phosphorylation of PLCgamma1. Permeabilization of RBL cells leads to loss of PLCgamma1 as well as PITP into the extracellular medium and this coincides with loss of antigen-stimulated lipid hydrolysis. Both PLCgamma1 and PITP were required to restore inositol lipid signaling. We conclude that (i) because the PI binding/transfer activities of PITP/SEC14p is the common feature shared by all three transfer proteins, it must be the relevant activity that determines their abilities to restore inositol lipid-mediated signaling and (ii) PITP is a general requirement for inositol lipid hydrolysis regardless of how and which isoform of PLC is activated by the appropriate agonist.
authors.affiliation: Department of Physiology, Rockfeller Building, University College London, United Kingdom.
authors.forename: E
authors.initials: E
authors.lastname: Cunningham
authors.forename: S K
authors.initials: SK
authors.lastname: Tan
authors.forename: P
authors.initials: P
authors.lastname: Swigart
authors.forename: J
authors.initials: J
authors.lastname: Hsuan
authors.forename: V
authors.initials: V
authors.lastname: Bankaitis
authors.forename: S
authors.initials: S
authors.lastname: Cockcroft
mesh_terms: D000818:Animals; D000941:Antigens / Q000494:pharmacology; D001426:Bacterial Proteins; D001483:Base Sequence; D002352:Carrier Proteins / Q000378:metabolism*; D002460:Cell Line; D017931:DNA Primers; D004789:Enzyme Activation; D019204:GTP-Binding Proteins / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D006868:Hydrolysis; D007527:Isoenzymes / Q000378:metabolism*; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010716:Phosphatidylinositols / Q000378:metabolism*; D051966:Phospholipase C gamma; D049294:Phospholipid Transfer Proteins; D051381:Rats; D012441:Saccharomyces cerevisiae / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins; D015398:Signal Transduction; D013301:Streptolysins / Q000494:pharmacology; D010738:Type C Phospholipases / Q000378:metabolism*
chemical_list: D000941:Antigens; D001426:Bacterial Proteins; D002352:Carrier Proteins; D017931:DNA Primers; D007527:Isoenzymes; D008565:Membrane Proteins; D010716:Phosphatidylinositols; D049294:Phospholipid Transfer Proteins; C118289:SEC24 protein, S cerevisiae; D029701:Saccharomyces cerevisiae Proteins; D013301:Streptolysins; C025964:streptolysin O; D010738:Type C Phospholipases; D051966:Phospholipase C gamma; D019204:GTP-Binding Proteins