1887

Abstract

We reported previously that Pin1 facilitates human immunodeficiency virus type 1 (HIV-1) uncoating by interacting with the capsid core through the phosphorylated Ser-Pro motif. However, the specific kinase responsible for Ser phosphorylation has remained unknown. Here, we showed that virion-associated extracellular signal-regulated kinase 2 (ERK2) phosphorylates Ser. The characterization of immature virions produced by exposing chronically HIV-1-infected CEM/LAV-1 cells to 10 µM saquinavir indicated that Ser is phosphorylated after the initiation of Pr55 processing. Furthermore, a mass spectrometry-based kinase assay demonstrated that ERK2 specifically phosphorylated the Ser residue in the Ser-Pro motif-containing substrate. The treatment of CEM/LAV-1 cells with the ERK2 inhibitor sc-222229 decreased the Ser phosphorylation level inside virions, and virus partially defective in Ser phosphorylation showed impaired reverse transcription and attenuated replication owing to attenuated Pin1-dependent uncoating. Furthermore, the suppression of ERK2 expression by RNA interference in CEM/LAV-1 cells resulted in suppressed ERK2 packaging inside virions and decreased the Ser phosphorylation level inside virions. Interestingly, the ERK2-packaging-defective virus showed impaired reverse transcription and attenuated HIV-1 replication. Taken together, these findings provide insights into the as-yet-obscure processes in Pin1-dependent HIV-1 uncoating.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.060053-0
2014-05-01
2020-01-25
Loading full text...

Full text loading...

/deliver/fulltext/jgv/95/5/1156.html?itemId=/content/journal/jgv/10.1099/vir.0.060053-0&mimeType=html&fmt=ahah

References

  1. Auewarakul P., Wacharapornin P., Srichatrapimuk S., Chutipongtanate S., Puthavathana P.. ( 2005; ). Uncoating of HIV-1 requires cellular activation. . Virology 337:, 93–101. [CrossRef] [PubMed]
    [Google Scholar]
  2. Bukrinskaya A. G., Ghorpade A., Heinzinger N. K., Smithgall T. E., Lewis R. E., Stevenson M.. ( 1996; ). Phosphorylation-dependent human immunodeficiency virus type 1 infection and nuclear targeting of viral DNA. . Proc Natl Acad Sci U S A 93:, 367–371. [CrossRef] [PubMed]
    [Google Scholar]
  3. Cartier C., Deckert M., Grangeasse C., Trauger R., Jensen F., Bernard A., Cozzone A., Desgranges C., Boyer V.. ( 1997; ). Association of ERK2 mitogen-activated protein kinase with human immunodeficiency virus particles. . J Virol 71:, 4832–4837.[PubMed]
    [Google Scholar]
  4. Cartier C., Hemonnot B., Gay B., Bardy M., Sanchiz C., Devaux C., Briant L.. ( 2003; ). Active cAMP-dependent protein kinase incorporated within highly purified HIV-1 particles is required for viral infectivity and interacts with viral capsid protein. . J Biol Chem 278:, 35211–35219. [CrossRef] [PubMed]
    [Google Scholar]
  5. Cen S., Khorchid A., Javanbakht H., Gabor J., Stello T., Shiba K., Musier-Forsyth K., Kleiman L.. ( 2001; ). Incorporation of lysyl-tRNA synthetase into human immunodeficiency virus type 1. . J Virol 75:, 5043–5048. [CrossRef] [PubMed]
    [Google Scholar]
  6. Derdeyn C. A., Decker J. M., Sfakianos J. N., Wu X., O’Brien W. A., Ratner L., Kappes J. C., Shaw G. M., Hunter E.. ( 2000; ). Sensitivity of human immunodeficiency virus type 1 to the fusion inhibitor T-20 is modulated by coreceptor specificity defined by the V3 loop of gp120. . J Virol 74:, 8358–8367. [CrossRef] [PubMed]
    [Google Scholar]
  7. Dismuke D. J., Aiken C.. ( 2006; ). Evidence for a functional link between uncoating of the human immunodeficiency virus type 1 core and nuclear import of the viral preintegration complex. . J Virol 80:, 3712–3720. [CrossRef] [PubMed]
    [Google Scholar]
  8. Fitzon T., Leschonsky B., Bieler K., Paulus C., Schröder J., Wolf H., Wagner R.. ( 2000; ). Proline residues in the HIV-1 NH2-terminal capsid domain: structure determinants for proper core assembly and subsequent steps of early replication. . Virology 268:, 294–307. [CrossRef] [PubMed]
    [Google Scholar]
  9. Forshey B. M., von Schwedler U., Sundquist W. I., Aiken C.. ( 2002; ). Formation of a human immunodeficiency virus type 1 core of optimal stability is crucial for viral replication. . J Virol 76:, 5667–5677. [CrossRef] [PubMed]
    [Google Scholar]
  10. Franke E. K., Yuan H. E., Luban J.. ( 1994; ). Specific incorporation of cyclophilin A into HIV-1 virions. . Nature 372:, 359–362. [CrossRef] [PubMed]
    [Google Scholar]
  11. Gabor J., Cen S., Javanbakht H., Niu M., Kleiman L.. ( 2002; ). Effect of altering the tRNA3 Lys concentration in human immunodeficiency virus type 1 upon its annealing to viral RNA, GagPol incorporation, and viral infectivity. . J Virol 76:, 9096–9102. [CrossRef] [PubMed]
    [Google Scholar]
  12. Gupta P., Singhal P. K., Rajendrakumar P., Padwad Y., Tendulkar A. V., Kalyanaraman V. S., Schmidt R. E., Srinivasan A., Mahalingam S.. ( 2011; ). Mechanism of host cell MAPK/ERK-2 incorporation into lentivirus particles: characterization of the interaction between MAPK/ERK-2 and proline-rich-domain containing capsid region of structural protein Gag. . J Mol Biol 410:, 681–697. [CrossRef] [PubMed]
    [Google Scholar]
  13. Hemonnot B., Cartier C., Gay B., Rebuffat S., Bardy M., Devaux C., Boyer V., Briant L.. ( 2004; ). The host cell MAP kinase ERK-2 regulates viral assembly and release by phosphorylating the p6gag protein of HIV-1. . J Biol Chem 279:, 32426–32434. [CrossRef] [PubMed]
    [Google Scholar]
  14. Hong W., Sano K., Morimatsu S., Scott D. R., Weeks D. L., Sachs G., Goto T., Mohan S., Harada F.. & other authors ( 2003; ). Medium pH-dependent redistribution of the urease of Helicobacter pylori . . J Med Microbiol 52:, 211–216. [CrossRef] [PubMed]
    [Google Scholar]
  15. Ikeda T., Nishitsuji H., Zhou X., Nara N., Ohashi T., Kannagi M., Masuda T.. ( 2004; ). Evaluation of the functional involvement of human immunodeficiency virus type 1 integrase in nuclear import of viral cDNA during acute infection. . J Virol 78:, 11563–11573. [CrossRef] [PubMed]
    [Google Scholar]
  16. Jacqué J. M., Mann A., Enslen H., Sharova N., Brichacek B., Davis R. J., Stevenson M.. ( 1998; ). Modulation of HIV-1 infectivity by MAPK, a virion-associated kinase. . EMBO J 17:, 2607–2618. [CrossRef] [PubMed]
    [Google Scholar]
  17. Kohno T., Fujioka Y., Goto T., Morimatsu S., Morita C., Nakano T., Sano K.. ( 1998; ). Contrast-enhancement for the image of human immunodeficiency virus from ultrathin section by immuno electron microscopy. . J Virol Methods 72:, 137–143. [CrossRef] [PubMed]
    [Google Scholar]
  18. Laemmli U. K.. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. . Nature 227:, 680–685. [CrossRef] [PubMed]
    [Google Scholar]
  19. Leschonsky B., Ludwig C., Bieler K., Wagner R.. ( 2007; ). Capsid stability and replication of human immunodeficiency virus type 1 are influenced critically by charge and size of Gag residue 183. . J Gen Virol 88:, 207–216. [CrossRef] [PubMed]
    [Google Scholar]
  20. Maréchal V., Clavel F., Heard J. M., Schwartz O.. ( 1998; ). Cytosolic Gag p24 as an index of productive entry of human immunodeficiency virus type 1. . J Virol 72:, 2208–2212.[PubMed]
    [Google Scholar]
  21. Misumi S., Inoue M., Dochi T., Kishimoto N., Hasegawa N., Takamune N., Shoji S.. ( 2010; ). Uncoating of human immunodeficiency virus type 1 requires prolyl isomerase Pin1. . J Biol Chem 285:, 25185–25195. [CrossRef] [PubMed]
    [Google Scholar]
  22. Müller B., Patschinsky T., Kräusslich H. G.. ( 2002; ). The late-domain-containing protein p6 is the predominant phosphoprotein of human immunodeficiency virus type 1 particles. . J Virol 76:, 1015–1024. [CrossRef] [PubMed]
    [Google Scholar]
  23. Ott D. E.. ( 1997; ). Cellular proteins in HIV virions. . Rev Med Virol 7:, 167–180. [CrossRef] [PubMed]
    [Google Scholar]
  24. Ott D. E.. ( 2009; ). Purification of HIV-1 virions by subtilisin digestion or CD45 immunoaffinity depletion for biochemical studies. . Methods Mol Biol 485:, 15–25. [CrossRef] [PubMed]
    [Google Scholar]
  25. Ott D. E., Coren L. V., Johnson D. G., Sowder R. C. II, Arthur L. O., Henderson L. E.. ( 1995; ). Analysis and localization of cyclophilin A found in the virions of human immunodeficiency virus type 1 MN strain. . AIDS Res Hum Retroviruses 11:, 1003–1006. [CrossRef] [PubMed]
    [Google Scholar]
  26. Rue S. M., Roos J. W., Tarwater P. M., Clements J. E., Barber S. A.. ( 2005; ). Phosphorylation and proteolytic cleavage of Gag proteins in budded simian immunodeficiency virus. . J Virol 79:, 2484–2492. [CrossRef] [PubMed]
    [Google Scholar]
  27. Thali M., Bukovsky A., Kondo E., Rosenwlrth B., Walsh C. T., Sodroski J., Göttlinger H. G.. ( 1994; ). Functional association of cyclophilin A with HIV-1 virions. . Nature 372:, 363–365. [CrossRef] [PubMed]
    [Google Scholar]
  28. Trubey C. M., Chertova E., Coren L. V., Hilburn J. M., Hixson C. V., Nagashima K., Lifson J. D., Ott D. E.. ( 2003; ). Quantitation of HLA class II protein incorporated into human immunodeficiency type 1 virions purified by anti-CD45 immunoaffinity depletion of microvesicles. . J Virol 77:, 12699–12709. [CrossRef] [PubMed]
    [Google Scholar]
  29. Yang X., Gabuzda D.. ( 1998; ). Mitogen-activated protein kinase phosphorylates and regulates the HIV-1 Vif protein. . J Biol Chem 273:, 29879–29887. [CrossRef] [PubMed]
    [Google Scholar]
  30. Yang X., Goncalves J., Gabuzda D.. ( 1996; ). Phosphorylation of Vif and its role in HIV-1 replication. . J Biol Chem 271:, 10121–10129. [CrossRef] [PubMed]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.060053-0
Loading
/content/journal/jgv/10.1099/vir.0.060053-0
Loading

Data & Media loading...

Most cited articles

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error