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Identification of interaction between HIV-1 glycoprotein 41 and integrase

Abstract

Human immunodeficiency virus-1 (HIV-1) encodes 15 viral proteins. Protein-protein interactions play a large role in the function of these proteins. In this study, we attempted to identify novel interactions between the HIV-1 proteins to better understand the role played by viral protein-protein interactions in the life cycle of HIV-1. Genes encoding the 15 viral proteins from the HIV-1 strain AD8 were inserted into the plasmids of a yeast two-hybrid system. By screening 120 pairs of proteins, interactions between seven pairs were found. This led to the discovery of an interaction between the HIV-1 proteins integrase (IN) and glycoprotein 41 (gp41), which was confirmed by both co-immunoprecipitation (Co-IP) assays and fluorescence resonance energy transfer (FRET) imaging in live cells. In addition, it was found that the amino acids at positions 76–100 of gp41 are required for it to bind to IN. Deletion of this region from gp41 prevented its interaction with IN and reduced the production of HIV-1 in 293T cells. This study provides new information on HIV-1 protein-protein interactions which improves the understanding of the biological functions of gp41 and IN during the virus life cycle.

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References

  • Archin NM, Sung JM, Garrido C, Soriano-Sarabia N, Margolis DM. 2014. Eradicating HIV-1 infection: seeking to clear a persistent pathogen. Nat Rev Microbiol, 12: 750–764.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Barre-Sinoussi F, Ross AL, Delfraissy JF. 2013. Past, present and future: 30 years of HIV research. Nat Rev Microbiol, 11: 877–883.

    CAS  Article  PubMed  Google Scholar 

  • Biswas N, Wang T, Ding M, Tumne A, Chen Y, Wang Q, Gupta P. 2012. ADAR1 is a novel multi targeted anti-HIV-1 cellular protein. Virology, 422: 265–277.

    CAS  Article  PubMed  Google Scholar 

  • Briggs JA, Simon MN, Gross I, Krausslich HG, Fuller SD, Vogt VM, Johnson MC. 2004. The stoichiometry of Gag protein in HIV-1. Nat Struct Mol Biol, 11: 672–675.

    CAS  Article  PubMed  Google Scholar 

  • Bukovsky A, Gottlinger H. 1996. Lack of integrase can markedly affect human immunodeficiency virus type 1 particle production in the presence of an active viral protease. J Virol, 70: 6820–6825.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Caffrey M. 2001. Model for the structure of the HIV gp41 ectodomain: insight into the intermolecular interactions of the gp41 loop. Biochim Biophys Acta, 31: 2–3.

    Google Scholar 

  • Caffrey M, Cai M, Kaufman J, Stahl SJ, Wingfield PT, Covell DG, Gronenborn AM, Clore GM. 1998. Three-dimensional solution structure of the 44 kDa ectodomain of SIV gp41. EMBO J, 17: 4572–4584.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Chan DC, Fass D, Berger JM, Kim PS. 1997. Core structure of gp41 from the HIV envelope glycoprotein. Cell, 89: 263–273.

    CAS  Article  PubMed  Google Scholar 

  • Chen JC, Krucinski J, Miercke LJ, Finer-Moore JS, Tang AH, Leavitt AD, Stroud RM. 2000. Crystal structure of the HIV-1 integrase catalytic core and C-terminal domains: a model for viral DNA binding. Proc Natl Acad Sci U S A, 97: 8233–8238.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Ciuffi A, Llano M, Poeschla E, Hoffmann C, Leipzig J, Shinn P, Ecker JR, Bushman F. 2005. A role for LEDGF/p75 in targeting HIV DNA integration. Nat Med, 11: 1287–1289.

    CAS  Article  PubMed  Google Scholar 

  • Cullen BR. 1998. HIV-1 auxiliary proteins: making connections in a dying cell. Cell, 93: 685–692.

    CAS  Article  PubMed  Google Scholar 

  • de Rocquigny H, Petitjean P, Tanchou V, Decimo D, Drouot L, Delaunay T, Darlix JL, Roques BP. 1997. The zinc fingers of HIV nucleocapsid protein NCp7 direct interactions with the viral regulatory protein Vpr. J Biol Chem, 272: 30753–30759.

    Article  PubMed  Google Scholar 

  • Frankel AD, Young JA. 1998. HIV-1: fifteen proteins and an RNA. Annu Rev Biochem, 67: 1–25.

    CAS  Article  PubMed  Google Scholar 

  • Freed EO. 2004. HIV-1 and the host cell: an intimate association. Trends Microbiol, 12: 170–177.

    CAS  Article  PubMed  Google Scholar 

  • Gleenberg IO, Herschhorn A, Hizi A. 2007. Inhibition of the activities of reverse transcriptase and integrase of human immunodeficiency virus type-1 by peptides derived from the homologous viral protein R (Vpr). J Mol Biol, 369: 1230–1243.

    CAS  Article  PubMed  Google Scholar 

  • Goff SP. 2007. Host factors exploited by retroviruses. Nat Rev Microbiol, 5: 253–263.

    CAS  Article  PubMed  Google Scholar 

  • Guarente L. 1983. Yeast promoters and lacZ fusions designed to study expression of cloned genes in yeast. Methods Enzymol, 101: 181–191.

    CAS  Article  PubMed  Google Scholar 

  • Hehl EA, Joshi P, Kalpana GV, Prasad VR. 2004. Interaction between human immunodeficiency virus type 1 reverse transcriptase and integrase proteins. J Virol, 78: 5056–5067.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Huang J, Wang F, Argyris E, Chen K, Liang Z, Tian H, Huang W, Squires K, Verlinghieri G, Zhang H. 2007. Cellular micro-RNAs contribute to HIV-1 latency in resting primary CD4+ T lymphocytes. Nat Med, 13: 1241–1247.

    CAS  Article  PubMed  Google Scholar 

  • Jacks T, Power MD, Masiarz FR, Luciw PA, Barr PJ, Varmus HE. 1988. Characterization of ribosomal frameshifting in HIV-1 gag-pol expression. Nature, 331: 280–283.

    CAS  Article  PubMed  Google Scholar 

  • Jenkins Y, Pornillos O, Rich RL, Myszka DG, Sundquist WI, Malim MH. 2001. Biochemical analyses of the interactions between human immunodeficiency virus type 1 Vpr and p6 (Gag). J Virol, 75: 10537–10542.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Karpova TS, Baumann CT, He L, Wu X, Grammer A, Lipsky P, Hager GL, McNally JG. 2003. Fluorescence resonance energy transfer from cyan to yellow fluorescent protein detected by acceptor photobleaching using confocal microscopy and a single laser. J Microsc, 209: 56–70.

    CAS  Article  PubMed  Google Scholar 

  • Kashanchi F, Piras G, Radonovich MF, Duvall JF, Fattaey A, Chiang CM, Roeder RG, Brady JN. 1994. Direct interaction of human TFIID with the HIV-1 transactivator tat. Nature, 367: 295–299.

    CAS  Article  PubMed  Google Scholar 

  • Konig R, Zhou Y, Elleder D, Diamond TL, Bonamy GM, Irelan JT, Chiang CY, Tu BP, De Jesus PD, Lilley CE, Seidel S, Opaluch AM, Caldwell JS, Weitzman MD, Kuhen KL, Bandyopadhyay S, Ideker T, Orth AP, Miraglia LJ, Bushman FD, Young JA, Chanda SK. 2008. Global analysis of host-pathogen interactions that regulate early-stage HIV-1 replication. Cell, 135: 49–60.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Levin A, Hayouka Z, Brack-Werner R, Volsky DJ, Friedler A, Loyter A. 2009. Novel regulation of HIV-1 replication and pathogenicity: Rev inhibition of integration. Protein Eng Des Sel, 22: 753–763.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Levy JA. 1993. Pathogenesis of human immunodeficiency virus infection. Microbiol Rev, 57: 183–289.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Marchio S, Alfano M, Primo L, Gramaglia D, Butini L, Gennero L, De Vivo E, Arap W, Giacca M, Pasqualini R, Bussolino F. 2005. Cell surface-associated Tat modulates HIV-1 infection and spreading through a specific interaction with gp120 viral envelope protein. Blood, 105: 2802–2811.

    CAS  Article  PubMed  Google Scholar 

  • Miyawaki A, Llopis J, Heim R, McCaffery JM, Adams JA, Ikura M, Tsien RY. 1997. Fluorescent indicators for Ca2+ based on green fluorescent proteins and calmodulin. Nature, 388: 882–887.

    CAS  Article  PubMed  Google Scholar 

  • Mockli N, Auerbach D. 2004. Quantitative beta-galactosidase assay suitable for high-throughput applications in the yeast twohybrid system. Biotechniques, 36: 872–876.

    PubMed  Google Scholar 

  • Murakami T, Freed EO. 2000. Genetic evidence for an interaction between human immunodeficiency virus type 1 matrix and alpha-helix 2 of the gp41 cytoplasmic tail. J Virol, 74: 3548–3554.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Oz I, Avidan O, Hizi A. 2002. Inhibition of the integrases of human immunodeficiency viruses type 1 and type 2 by reverse transcriptases. Biochem J, 361: 557–566.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Piot P, Abdool Karim SS, Hecht R, Legido-Quigley H, Buse K, Stover J, Resch S, Ryckman T, Mogedal S, Dybul M, Goosby E, Watts C, Kilonzo N, McManus J, Sidibe M. 2015. Defeating AIDS—advancing global health. Lancet, 386: 171–218.

    Article  PubMed  Google Scholar 

  • Quinn TC. 2008. HIV epidemiology and the effects of antiviral therapy on long-term consequences. AIDS, 22, Suppl 3:S7–S 12.

    Google Scholar 

  • Roche J, Louis JM, Grishaev A, Ying J, Bax A. 2014. Dissociation of the trimeric gp41 ectodomain at the lipid-water interface suggests an active role in HIV-1 Env-mediated membrane fusion. Proc Natl Acad Sci U S A, 111: 3425–3430.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Rosenbluh J, Hayouka Z, Loya S, Levin A, Armon-Omer A, Britan E, Hizi A, Kotler M, Friedler A, Loyter A. 2007. Interaction between HIV-1 Rev and integrase proteins: a basis for the development of anti-HIV peptides. J Biol Chem, 282: 15743–15753.

    CAS  Article  PubMed  Google Scholar 

  • Selig L, Pages JC, Tanchou V, Preveral S, Berlioz-Torrent C, Liu LX, Erdtmann L, Darlix J, Benarous R, Benichou S. 1999. Interaction with the p6 domain of the gag precursor mediates incorporation into virions of Vpr and Vpx proteins from primate lentiviruses. J Virol, 73: 592–600.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Shin CG, Taddeo B, Haseltine WA, Farnet CM. 1994. Genetic analysis of the human immunodeficiency virus type 1 integrase protein. J Virol, 68: 1633–1642.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Suzuki S, Urano E, Hashimoto C, Tsutsumi H, Nakahara T, Tanaka T, Nakanishi Y, Maddali K, Han Y, Hamatake M, Miyauchi K, Pommier Y, Beutler JA, Sugiura W, Fuji H, Hoshino T, Itotani K, Nomura W, Narumi T, Yamamoto N, Komano JA, Tamamura H. 2010. Peptide HIV-1 integrase inhibitors from HIV-1 gene products. J Med Chem, 53: 5356–5360.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Swanson CM, Malim MH. 2008. SnapShot: HIV-1 proteins. Cell, 133: 742, 742.e1.

  • Tang H, Kuhen KL, Wong-Staal F. 1999. Lentivirus replication and regulation. Annu Rev Genet, 33: 133–170.

    CAS  Article  PubMed  Google Scholar 

  • Tavassoli A. 2011. Targeting the protein-protein interactions of the HIV lifecycle. Chem Soc Rev, 40: 1337–1346.

    CAS  Article  PubMed  Google Scholar 

  • Tekeste SS, Wilkinson TA, Weiner EM, Xu X, Miller JT, Le Grice SF, Clubb RT, Chow SA. 2015. Interaction between Reverse Transcriptase and Integrase Is Required for Reverse Transcription during HIV-1 Replication. J Virol, 89: 12058–12069.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Wilkinson TA, Januszyk K, Phillips ML, Tekeste SS, Zhang M, Miller JT, Le Grice SF, Clubb RT, Chow SA. 2009. Identifying and characterizing a functional HIV-1 reverse transcriptasebinding site on integrase. J Biol Chem, 284: 7931–7939.

    CAS  Article  PubMed  PubMed Central  Google Scholar 

  • Wu L, Gerard NP, Wyatt R, Choe H, Parolin C, Ruffing N, Borsetti A, Cardoso AA, Desjardin E, Newman W, Gerard C, Sodroski J. 1996. CD4-induced interaction of primary HIV-1 gp120 glycoproteins with the chemokine receptor CCR-5. Nature, 384: 179–183.

    CAS  Article  PubMed  Google Scholar 

  • Wyatt R, Desjardin E, Olshevsky U, Nixon C, Binley J, Olshevsky V, Sodroski J. 1997. Analysis of the interaction of the human immunodeficiency virus type 1 gp120 envelope glycoprotein with the gp41 transmembrane glycoprotein. J Virol, 71: 9722–9731.

    CAS  PubMed  PubMed Central  Google Scholar 

  • Wyatt R, Sodroski J. 1998. The HIV-1 envelope glycoproteins: fusogens, antigens, and immunogens. Science, 280: 1884–1888.

    CAS  Article  PubMed  Google Scholar 

  • Yung E, Sorin M, Pal A, Craig E, Morozov A, Delattre O, Kappes J, Ott D, Kalpana GV. 2001. Inhibition of HIV-1 virion production by a transdominant mutant of integrase interactor 1. Nat Med, 7: 920–926.

    CAS  Article  PubMed  Google Scholar 

  • Zhou H, Xu M, Huang Q, Gates AT, Zhang XD, Castle JC, Stec E, Ferrer M, Strulovici B, Hazuda DJ, Espeseth AS. 2008. Genome-scale RNAi screen for host factors required for HIV replication. Cell Host Microbe, 4: 495–504.

    CAS  Article  PubMed  Google Scholar 

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Correspondence to Xian-En Zhang or Zongqiang Cui.

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These authors contributed equally to this work.

ORCID: 0000-0003-1564-272X

ORCID: 0000-0002-0000-148X

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Zhang, X., Zhang, F., Ma, X. et al. Identification of interaction between HIV-1 glycoprotein 41 and integrase. Virol. Sin. 31, 415–424 (2016). https://doi.org/10.1007/s12250-016-3820-7

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  • DOI: https://doi.org/10.1007/s12250-016-3820-7

Keywords

  • human immunodeficiency virus-1 (HIV-1)
  • glycoprotein 41 (gp41)
  • integrase (IN)
  • protein-protein interactions
  • yeast two-hybrid assay