1887

Journal of General Virology header logo

Volume 89, Issue 12

Substitution of the myristoylation signal of human immunodeficiency virus type 1 Pr55 with the phospholipase C-1 pleckstrin homology domain results in infectious pseudovirion production Free

Abstract

The matrix domain (MA) of human immunodeficiency virus type 1 Pr55 is covalently modified with a myristoyl group that mediates efficient viral production. However, the role of myristoylation, particularly in the viral entry process, remains uninvestigated. This study replaced the myristoylation signal of MA with a well-studied phosphatidylinositol 4,5-biphosphate-binding plasma membrane (PM) targeting motif, the phospholipase C-1 pleckstrin homology (PH) domain. PH–Gag–Pol PM targeting and viral production efficiencies were improved compared with Gag–Pol, consistent with the estimated increases in Gag–PM affinity. Both virions were recovered in similar sucrose density-gradient fractions and had similar mature virion morphologies. Importantly, PH–Gag–Pol and Gag–Pol pseudovirions had almost identical infectivity, suggesting a dispensable role for myristoylation in the virus life cycle. PH–Gag–Pol might be useful in separating the myristoylation-dependent processes from the myristoylation-independent processes. This the first report demonstrating infectious pseudovirion production without myristoylated Pr55.

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2008-12-01
2024-04-28
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References

  1. Berridge M. J. 1993; Inositol trisphosphate and calcium signalling. Nature 361:315–325 [CrossRef]
     [Google Scholar]
  2. Bryant M., Ratner L. 1990; Myristoylation-dependent replication and assembly of human immunodeficiency virus 1. Proc Natl Acad Sci U S A 87:523–527 [CrossRef]
     [Google Scholar]
  3. Bukrinskaya A. 2007; HIV-1 matrix protein: a mysterious regulator of the viral life cycle. Virus Res 124:1–11 [CrossRef]
     [Google Scholar]
  4. de Oliveira T., Engelbrecht S., , Janse van Rensburg E., Gordon M., Bishop K., zur Megede J., Barnett S. W., Cassol S. 2003; Variability at human immunodeficiency virus type 1 subtype C protease cleavage sites: an indication of viral fitness?. J Virol 77:9422–9430 [CrossRef]
     [Google Scholar]
  5. Ferguson K. M., Lemmon M. A., Schlessinger J., Sigler P. B. 1995a; Structure of the high affinity complex of inositol trisphosphate with a phospholipase C pleckstrin homology domain. Cell 83:1037–1046 [CrossRef]
     [Google Scholar]
  6. Ferguson K. M., Lemmon M. A., Sigler P. B., Schlessinger J. 1995b; Scratching the surface with the PH domain. Nat Struct Biol 2:715–718 [CrossRef]
     [Google Scholar]
  7. Fiorentini S., Marini E., Caracciolo S., Caruso A. 2006; Functions of the HIV-1 matrix protein p17. New Microbiol 29:1–10
     [Google Scholar]
  8. Futahashi Y., Komano J., Urano E., Aoki T., Hamatake M., Miyauchi K., Yoshida T., Koyanagi Y., Matsuda Z., Yamamoto N. 2007; Separate elements are required for ligand-dependent and -independent internalization of metastatic potentiator CXCR4. Cancer Sci 98:373–379 [CrossRef]
     [Google Scholar]
  9. Göttlinger H. G., Sodroski J. G., Haseltine W. A. 1989; Role of capsid precursor processing and myristoylation in morphogenesis and infectivity of human immunodeficiency virus type 1. Proc Natl Acad Sci U S A 86:5781–5785 [CrossRef]
     [Google Scholar]
  10. Harlan J. E., Hajduk P. J., Yoon H. S., Fesik S. W. 1994; Pleckstrin homology domains bind to phosphatidylinositol-4,5-bisphosphate. Nature 371:168–170 [CrossRef]
     [Google Scholar]
  11. Harlan J. E., Yoon H. S., Hajduk P. J., Fesik S. W. 1995; Structural characterization of the interaction between a pleckstrin homology domain and phosphatidylinositol 4,5-bisphosphate. Biochemistry 34:9859–9864 [CrossRef]
     [Google Scholar]
  12. Hearps A. C., Jans D. A. 2007; Regulating the functions of the HIV-1 matrix protein. AIDS Res Hum Retroviruses 23:341–346 [CrossRef]
     [Google Scholar]
  13. Jouvenet N., Neil S. J., Bess C., Johnson M. C., Virgen C. A., Simon S. M., Bieniasz P. D. 2006; Plasma membrane is the site of productive HIV-1 particle assembly. PLoS Biol 4:e435 [CrossRef]
     [Google Scholar]
  14. Kiernan R. E., Ono A., Englund G., Freed E. O. 1998; Role of matrix in an early postentry step in the human immunodeficiency virus type 1 life cycle. J Virol 72:4116–4126
     [Google Scholar]
  15. Klein K. C., Reed J. C., Lingappa J. R. 2007; Intracellular destinies: degradation, targeting, assembly, and endocytosis of HIV Gag. AIDS Rev 9:150–161
     [Google Scholar]
  16. Komano J., Miyauchi K., Matsuda Z., Yamamoto N. 2004; Inhibiting the Arp2/3 complex limits infection of both intracellular mature vaccinia virus and primate lentiviruses. Mol Biol Cell 15:5197–5207 [CrossRef]
     [Google Scholar]
  17. Lemmon M. A., Ferguson K. M., O'Brien R., Sigler P. B., Schlessinger J. 1995; Specific and high-affinity binding of inositol phosphates to an isolated pleckstrin homology domain. Proc Natl Acad Sci U S A 92:10472–10476 [CrossRef]
     [Google Scholar]
  18. Ono A., Ablan S. D., Lockett S. J., Nagashima K., Freed E. O. 2004; Phosphatidylinositol (4,5) bisphosphate regulates HIV-1 Gag targeting to the plasma membrane. Proc Natl Acad Sci U S A 101:14889–14894 [CrossRef]
     [Google Scholar]
  19. Pal R., Reitz M. S. Jr, Tschachler E., Gallo R. C., Sarngadharan M. G., Veronese F. D. 1990; Myristoylation of gag proteins of HIV-1 plays an important role in virus assembly. AIDS Res Hum Retroviruses 6:721–730 [CrossRef]
     [Google Scholar]
  20. Provitera P., El-Maghrabi R., Scarlata S. 2006; The effect of HIV-1 Gag myristoylation on membrane binding. Biophys Chem 119:23–32 [CrossRef]
     [Google Scholar]
  21. Rhee S. G. 2001; Regulation of phosphoinositide-specific phospholipase C. Annu Rev Biochem 70:281–312 [CrossRef]
     [Google Scholar]
  22. Saad J. S., Miller J., Tai J., Kim A., Ghanam R. H., Summers M. F. 2006; Structural basis for targeting HIV-1 Gag proteins to the plasma membrane for virus assembly. Proc Natl Acad Sci U S A 103:11364–11369 [CrossRef]
     [Google Scholar]
  23. Scholz I., Still A., Dhenub T. C., Coday K., Webb M., Barklis E. 2008; Analysis of human immunodeficiency virus matrix domain replacements. Virology 371:322–335 [CrossRef]
     [Google Scholar]
  24. Stauffer T. P., Ahn S., Meyer T. 1998; Receptor-induced transient reduction in plasma membrane PtdIns(4,5)P2 concentration monitored in living cells. Curr Biol 8:343–346 [CrossRef]
     [Google Scholar]
  25. Swanstrom R., Wills J. W. 1997; Synthesis, assembly, and processing of viral proteins. In Retroviruses pp 263–334Edited by Coffin J. M., Hughes S. H., Varmus. Cold Spring Harbor: Cold Spring Harbor Laboratory Press;
     [Google Scholar]
  26. Tall E. G., Spector I., Pentyala S. N., Bitter I., Rebecchi M. J. 2000; Dynamics of phosphatidylinositol 4,5-bisphosphate in actin-rich structures. Curr Biol 10:743–746 [CrossRef]
     [Google Scholar]
  27. Wagner R., Graf M., Bieler K., Wolf H., Grunwald T., Foley P., Uberla K. 2000; Rev-independent expression of synthetic gag-pol genes of human immunodeficiency virus type 1 and simian immunodeficiency virus: implications for the safety of lentiviral vectors. Hum Gene Ther 11:2403–2413 [CrossRef]
     [Google Scholar]
  28. Wang C. T., Zhang Y., McDermott J., Barklis E. 1993; Conditional infectivity of a human immunodeficiency virus matrix domain deletion mutant. J Virol 67:7067–7076
     [Google Scholar]
  29. Yagisawa H., Hirata M., Kanematsu T., Watanabe Y., Ozaki S., Sakuma K., Tanaka H., Yabuta N., Kamata H. other authors 1994; Expression and characterization of an inositol 1,4,5-trisphosphate binding domain of phosphatidylinositol-specific phospholipase C- δ 1. J Biol Chem 269:20179–20188
     [Google Scholar]
  30. Zhou W., Parent L. J., Wills J. W., Resh M. D. 1994; Identification of a membrane-binding domain within the amino-terminal region of human immunodeficiency virus type 1 Gag protein which interacts with acidic phospholipids. J Virol 68:2556–2569
     [Google Scholar]
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Substitution of the myristoylation signal of human immunodeficiency virus type 1 Pr55Gag with the phospholipase C-δ1 pleckstrin homology domain results in infectious pseudovirion production
J. Gen. Virol. 89, 3144 (2008); https://doi.org/10.1099/vir.0.2008/004820-0
/content/journal/jgv/10.1099/vir.0.2008/004820-0
/content/journal/jgv/10.1099/vir.0.2008/004820-0
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