Involvement of a herbimycin A-sensitive protein tyrosine kinase in extracellular action of HIV-1 Nef.
Extracellular Nef which has been implicated in disease progression and development of AIDS induces IL-10, a potent immunosuppressive cytokine, in vitro. The present study was designed to examine whether the action of extracellular Nef is a protein tyrosine kinase (PTK)-dependent event. Anti-phosphotyrosine immunoblotting reveals that recombinant HIV-1 Nef induces rapid tyrosyl phosphorylation of several cellular proteins in human peripheral blood mononuclear cells. Pre-treatment of cells with herbimycin A, but not with genistein, significantly abolishes the Nef-induced tyrosine phosphorylation of cellular proteins. Furthermore, ELISA and RNase protection assays show that herbimycin A significantly blocks Nef-induced production of IL-10 at both the protein and the mRNA level. Genistein and aminogenistein have a much less blocking effect on the ability of Nef to induce IL-10. These results provide evidence for the involvement of a herbimycin A-sensitive PTK in the signal transduction pathway for exogenous HIV-1 Nef.
Haraguchi
S
S
Department of Pediatrics, University of South Florida College of Medicine, All Children's Hospital, 801 Sixth Street South, St. Petersburg, FL 33701, USA.
Brigino-Buenaventura
E N
EN
Hitchcock
R
R
James-Yarish
M
M
Good
R A
RA
Day
N K
NK
D016227:Benzoquinones; D002478:Cells, Cultured; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005110:Extracellular Space / Q000378:metabolism; D015693:Gene Products, nef / Q000378:metabolism* / Q000494:pharmacology; D019833:Genistein / Q000494:pharmacology; D015497:HIV-1 / Q000378:metabolism*; D006801:Humans; D016753:Interleukin-10 / Q000096:biosynthesis / Q000235:genetics; D047029:Lactams, Macrocyclic; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors* / Q000378:metabolism*; D011809:Quinones / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology; D017828:Rifabutin / Q000031:analogs & derivatives; D015398:Signal Transduction; D054311:nef Gene Products, Human Immunodeficiency Virus
D016227:Benzoquinones; D004791:Enzyme Inhibitors; D015693:Gene Products, nef; D047029:Lactams, Macrocyclic; D011809:Quinones; D012333:RNA, Messenger; D011994:Recombinant Proteins; D054311:nef Gene Products, Human Immunodeficiency Virus; D016753:Interleukin-10; D017828:Rifabutin; C020754:herbimycin; D019833:Genistein; D011505:Protein-Tyrosine Kinases