Protein kinase C-mediated phosphorylation of HIV-I nef in human cell lines - PubMed (original) (raw)
. 1997 May 9;272(19):12289-94.
doi: 10.1074/jbc.272.19.12289.
Affiliations
- PMID: 9139671
- DOI: 10.1074/jbc.272.19.12289
Free article
Protein kinase C-mediated phosphorylation of HIV-I nef in human cell lines
K Coates et al. J Biol Chem. 1997.
Free article
Abstract
Stable human cell lines expressing the human immunodeficiency virus type I (HIV-I) Nef protein from inducible promoters were used to analyze the phosphorylation status of Nef in vivo. Nef phosphorylation in both HeLa and Jurkat cells was stimulated by phorbol ester treatment. Phosphoamino acid analysis revealed a predominance of phosphoserine with a small proportion of phosphothreonine. Treatment of cells with selective protein kinase inhibitors revealed that Nef phosphorylation was markedly reduced by bisindolylmaleimide, an inhibitor of protein kinase C, but was unaffected by inhibitors of mitogen-activated protein kinase kinase or cAMP-dependent kinase. These data implicate protein kinase C in Nef phosphorylation in vivo, and thus confirm and extend earlier in vitro data. Phosphorylation of a nonmyristoylated Nef mutant was impaired, suggesting that membrane targeting of Nef was required for phosphorylation. This was expected given that activated protein kinase C translocates from the cytosol to the plasma membrane. However, analysis of the subcellular localization of phosphorylated wild-type Nef revealed that both the cytosolic and membrane-associated pools of Nef were phosphorylated to an equivalent extent. Thus the significance of myristoylation for Nef function may be in influencing protein conformation, although these data could be explained by a transient and dynamic interaction between myristoylated Nef and the plasma membrane.
Similar articles
- The human immunodeficiency virus type 1 Nef protein functions as a protein kinase C substrate in vitro.
Coates K, Harris M. Coates K, et al. J Gen Virol. 1995 Apr;76 ( Pt 4):837-44. doi: 10.1099/0022-1317-76-4-837. J Gen Virol. 1995. PMID: 9049329 - Induction of phosphorylation of human immunodeficiency virus type 1 Nef and enhancement of CD4 downregulation by phorbol myristate acetate.
Luo T, Downing JR, Garcia JV. Luo T, et al. J Virol. 1997 Mar;71(3):2535-9. doi: 10.1128/JVI.71.3.2535-2539.1997. J Virol. 1997. PMID: 9032396 Free PMC article. - Cellular distribution of HIV type 1 Nef protein: identification of domains in Nef required for association with membrane and detergent-insoluble cellular matrix.
Kaminchik J, Margalit R, Yaish S, Drummer H, Amit B, Sarver N, Gorecki M, Panet A. Kaminchik J, et al. AIDS Res Hum Retroviruses. 1994 Aug;10(8):1003-10. doi: 10.1089/aid.1994.10.1003. AIDS Res Hum Retroviruses. 1994. PMID: 7811531 - In vitro binding and phosphorylation of human immunodeficiency virus type 1 Nef protein by serine/threonine protein kinase.
Bodéus M, Marie-Cardine A, Bougeret C, Ramos-Morales F, Benarous R. Bodéus M, et al. J Gen Virol. 1995 Jun;76 ( Pt 6):1337-44. doi: 10.1099/0022-1317-76-6-1337. J Gen Virol. 1995. PMID: 7540194 - Production of a non-functional nef protein in human immunodeficiency virus type 1-infected CEM cells.
Laurent AG, Hovanessian AG, Rivière Y, Krust B, Regnault A, Montagnier L, Findeli A, Kieny MP, Guy B. Laurent AG, et al. J Gen Virol. 1990 Oct;71 ( Pt 10):2273-81. doi: 10.1099/0022-1317-71-10-2273. J Gen Virol. 1990. PMID: 1977871
Cited by
- How HIV Nef Proteins Hijack Membrane Traffic To Promote Infection.
Buffalo CZ, Iwamoto Y, Hurley JH, Ren X. Buffalo CZ, et al. J Virol. 2019 Nov 26;93(24):e01322-19. doi: 10.1128/JVI.01322-19. Print 2019 Dec 15. J Virol. 2019. PMID: 31578291 Free PMC article. Review. - HIV-1 Nef and Vpu are functionally redundant broad-spectrum modulators of cell surface receptors, including tetraspanins.
Haller C, Müller B, Fritz JV, Lamas-Murua M, Stolp B, Pujol FM, Keppler OT, Fackler OT. Haller C, et al. J Virol. 2014 Dec;88(24):14241-57. doi: 10.1128/JVI.02333-14. Epub 2014 Oct 1. J Virol. 2014. PMID: 25275127 Free PMC article. - Heterologous Src homology 4 domains support membrane anchoring and biological activity of HIV-1 Nef.
Geist MM, Pan X, Bender S, Bartenschlager R, Nickel W, Fackler OT. Geist MM, et al. J Biol Chem. 2014 May 16;289(20):14030-44. doi: 10.1074/jbc.M114.563528. Epub 2014 Apr 4. J Biol Chem. 2014. PMID: 24706755 Free PMC article. - Conformational transition of membrane-associated terminally acylated HIV-1 Nef.
Akgun B, Satija S, Nanda H, Pirrone GF, Shi X, Engen JR, Kent MS. Akgun B, et al. Structure. 2013 Oct 8;21(10):1822-33. doi: 10.1016/j.str.2013.08.008. Epub 2013 Sep 12. Structure. 2013. PMID: 24035710 Free PMC article. - Neutron reflectometry study of the conformation of HIV Nef bound to lipid membranes.
Kent MS, Murton JK, Sasaki DY, Satija S, Akgun B, Nanda H, Curtis JE, Majewski J, Morgan CR, Engen JR. Kent MS, et al. Biophys J. 2010 Sep 22;99(6):1940-8. doi: 10.1016/j.bpj.2010.07.016. Biophys J. 2010. PMID: 20858440 Free PMC article.