title: ESCRT machinery potentiates HIV-1 utilization of the PI(4,5)P(2)-PLC-IP3R-Ca(2+) signaling cascade.
abstract: Human immunodeficiency virus type 1 (HIV-1) release efficiency is directed by late (L) domain motifs in the viral structural precursor polyprotein Gag, which serve as links to the ESCRT (endosomal sorting complex required for transport) machinery. Linkage is normally through binding of Tsg101, an ESCRT-1 component, to the P(7)TAP motif in the p6 region of Gag. In its absence, budding is directed by binding of Alix, an ESCRT adaptor protein, to the LY(36)PX(n)L motif in Gag. We recently showed that budding requires activation of the inositol 1,4,5-triphosphate receptor (IP3R), a protein that "gates" Ca(2+) release from intracellular stores, triggers Ca(2+) cell influx and thereby functions as a major regulator of Ca(2+) signaling. In the present study, we determined whether the L domain links Gag to Ca(2+) signaling machinery. Depletion of IP3R and inactivation of phospholipase C (PLC) inhibited budding whether or not Tsg101 was bound to Gag. PLC hydrolysis of phosphatidylinositol-(4,5)-bisphosphate generates inositol (1,4,5)-triphosphate, the ligand that activates IP3R. However, with Tsg101 bound, Gag release was independent of Gq-mediated activation of PLC, and budding was readily enhanced by pharmacological stimulation of PLC. Moreover, IP3R was redistributed to the cell periphery and cytosolic Ca(2+) was elevated, events indicative of induction of Ca(2+) signaling. The results suggest that L domain function, ESCRT machinery and Ca(2+) signaling are linked events in Gag release.
authors.affiliation: Department of Molecular Genetics and Microbiology, Stony Brook University, Stony Brook, NY 11794-5222, USA.
authors.forename: Lorna S
authors.initials: LS
authors.lastname: Ehrlich
authors.forename: Gisselle N
authors.initials: GN
authors.lastname: Medina
authors.forename: Carol A
authors.initials: CA
authors.lastname: Carter
mesh_terms: D000818:Animals; D015153:Blotting, Western; D019556:COS Cells; D002122:Calcium Chloride / Q000494:pharmacology; D020013:Calcium Signaling; D002462:Cell Membrane / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D056827:Endosomal Sorting Complexes Required for Transport / Q000378:metabolism*; D015658:HIV Infections / Q000378:metabolism / Q000821:virology; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D053496:Inositol 1,4,5-Trisphosphate Receptors / Q000378:metabolism*; D015759:Ionomycin / Q000494:pharmacology; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000378:metabolism*; D014157:Transcription Factors / Q000378:metabolism; D010738:Type C Phospholipases / Q000378:metabolism*; D054301:gag Gene Products, Human Immunodeficiency Virus / Q000378:metabolism*; D000099177:Tsg101 Protein
chemical_list: D002122:Calcium Chloride; D004268:DNA-Binding Proteins; D056827:Endosomal Sorting Complexes Required for Transport; D053496:Inositol 1,4,5-Trisphosphate Receptors; D015759:Ionomycin; D019269:Phosphatidylinositol 4,5-Diphosphate; D014157:Transcription Factors; D010738:Type C Phospholipases; D054301:gag Gene Products, Human Immunodeficiency Virus; D000099177:Tsg101 Protein