title: Nuclear phosphoinositide-specific phospholipase C β1 controls cytoplasmic CCL2 mRNA levels in HIV-1 gp120-stimulated primary human macrophages.
abstract: HIV-1 envelope glycoprotein gp120 induces, independently of infection, the release of CCL2 from macrophages. In turn, this chemokine acts as an autocrine factor enhancing viral replication. In this study, we show for the first time that phosphoinositide-specific phospholipase C (PI-PLC) is required for the production of CCL2 triggered by gp120 in macrophages. Using a combination of confocal laser-scanner microscopy, pharmacologic inhibition, western blotting and fluorescence-activated cell sorter analysis, we demonstrate that gp120 interaction with CCR5 leads to nuclear localization of the PI-PLC β1 isozyme mediated by mitogen-activated protein kinase ERK-1/2. Notably, phosphatidylcholine-specific phospholipase C (PC-PLC), previously reported to be required for NF-kB-mediated CCL2 production induced by gp120 in macrophages, drives both ERK1/2 activation and PI-PLC β1 nuclear localization induced by gp120. PI-PLC β1 activation through CCR5 is also triggered by the natural chemokine ligand CCL4, but independently of ERK1/2. Finally, PI-PLC inhibition neither blocks gp120-mediated NF-kB activation nor overall accumulation of CCL2 mRNA, whereas it decreases CCL2 transcript level in the cytoplasm. These results identify nuclear PI-PLC β1 as a new intermediate in the gp120-triggered PC-PLC-driven signal transduction pathway leading to CCL2 secretion in macrophages. The finding that a concerted gp120-mediated signaling involving both PC- and PI-specific PLCs is required for the expression of CCL2 in macrophages suggests that this signal transduction pathway may also be relevant for the modulation of viral replication in these cells. Thus, this study may contribute to identify novel targets for therapeutic intervention in HIV-1 infection.
authors.affiliation: Department of Hematology, Oncology and Molecular Medicine, Istituto Superiore di Sanità, Rome, Italy.
authors.forename: Francesca
authors.initials: F
authors.lastname: Spadaro
authors.forename: Serena
authors.initials: S
authors.lastname: Cecchetti
authors.forename: Cristina
authors.initials: C
authors.lastname: Purificato
authors.forename: Michela
authors.initials: M
authors.lastname: Sabbatucci
authors.forename: Franca
authors.initials: F
authors.lastname: Podo
authors.forename: Carlo
authors.initials: C
authors.lastname: Ramoni
authors.forename: Sandra
authors.initials: S
authors.lastname: Gessani
authors.forename: Laura
authors.initials: L
authors.lastname: Fantuzzi
mesh_terms: D002070:Butadienes / Q000494:pharmacology; D002467:Cell Nucleus / Q000378:metabolism; D002469:Cell Separation; D018932:Chemokine CCL2 / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism*; D005434:Flow Cytometry; D015699:HIV Envelope Protein gp120 / Q000378:metabolism*; D015497:HIV-1; D006801:Humans; D020935:MAP Kinase Signaling System; D008264:Macrophages / Q000378:metabolism / Q000821:virology*; D018613:Microscopy, Confocal; D009000:Monocytes / Q000166:cytology; D009570:Nitriles / Q000494:pharmacology; D010726:Phosphodiesterase Inhibitors / Q000494:pharmacology; D054799:Phospholipase C beta / Q000378:metabolism*; D010742:Phospholipid Ethers / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism
chemical_list: D002070:Butadienes; C111835:CCL2 protein, human; D018932:Chemokine CCL2; D015699:HIV Envelope Protein gp120; D009570:Nitriles; D010726:Phosphodiesterase Inhibitors; D010742:Phospholipid Ethers; D012333:RNA, Messenger; D011994:Recombinant Proteins; C113580:U 0126; C026659:edelfosine; D054799:Phospholipase C beta
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