CCR5 signaling through phospholipase D involves p44/42 MAP-kinases and promotes HIV-1 LTR-directed gene expression.
The chemokine receptor CCR5 plays an important role as an entry gate for the human immunodeficiency virus-1 (HIV-1) and for viral postentry events. Among signal transducers used by chemoattractant receptors, the phosphatidylcholine-specific phospholipase D (PLD) produces large amounts of second messengers in most cell types. However, the relevance of PLD isoforms to CCR5 signaling and HIV-1 infection process remains unexplored. We show here that CCR5 activation by MIP-1beta in HeLa-MAGI cells triggered a rapid and substantial PLD activity, as assessed by mass choline production. This activity required the activation of ERK1/2-MAP kinases and involved both PLD1 and PLD2. MIP-1beta also promoted the activation of an HIV-1 long terminal repeat (LTR) by the transactivator Tat in HeLa P4.2 cells through a process involving ERK1/2. Expression of wild-type and catalytically inactive PLDs dramatically boosted and inhibited the LTR activation, respectively, without altering Tat expression. Wild-type and inactive PLDs also respectively potentiated and inhibited HIV-1(BAL) replication in MAGI cells. Finally, in monocytic THP-1 cells, antisense oligonucleotides to both PLDs dramatically inhibited the HIV-1 replication. Thus, PLD is activated downstream of ERK1/2 upon CCR5 activation and plays a major role in promoting HIV-1 LTR transactivation and virus replication, which may open novel perspectives to anti-HIV-1 strategies.
Paruch
Sylvain
S
Institut Cochin, Université Paris Descartes, CNRS (UMR 8104), Paris, France.
Heinis
Mylène
M
Lemay
Julie
J
Hoeffel
Guillaume
G
Marañón
Concepción
C
Hosmalin
Anne
A
Périanin
Axel
A
D002794:Choline / Q000096:biosynthesis; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D016325:HIV Long Terminal Repeat / Q000502:physiology*; D015497:HIV-1 / Q000201:enzymology / Q000235:genetics* / Q000502:physiology; D006367:HeLa Cells; D006801:Humans; D020935:MAP Kinase Signaling System / Q000502:physiology*; D019950:Mitogen-Activated Protein Kinase 1 / Q000502:physiology*; D048052:Mitogen-Activated Protein Kinase 3 / Q000502:physiology*; D010739:Phospholipase D / Q000378:metabolism / Q000502:physiology*; D019713:Receptors, CCR5 / Q000378:metabolism / Q000502:physiology*; D015398:Signal Transduction / Q000502:physiology; D015533:Transcriptional Activation / Q000235:genetics; D014779:Virus Replication / Q000502:physiology
D019713:Receptors, CCR5; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D010739:Phospholipase D; D002794:Choline