Host Cofactors and Pharmacologic Ligands Share an Essential Interface in HIV-1 Capsid That Is Lost upon Disassembly

English version České info

The early steps of HIV-1 infection are poorly understood, in part because of the difficulty in obtaining high-resolution information on encapsidated virus and its interaction with host cofactors. This, in turn, has made it difficult to design effective anti-capsid (CA) drugs. In our present study, we have used stabilized hexamers of HIV-1 CA to obtain complexed crystal structures with two cellular cofactors that are important for HIV-1 infection. These structures and accompanying virology reveal an essential interface in the capsid of HIV-1 that is lost upon viral uncoating. This interface is used to recruit both the nuclear targeting cofactor CPSF6 and NUP153, a nuclear pore component that facilitates nuclear entry. The high-resolution information provided by these structures reveals that the interface is degenerate and CA mutations can be made that selectively perturb sensitivity to each cofactor. This interface is also competed by two antiviral drugs, PF74 and BI-2, whose different mechanisms of action are not fully understood. We show that PF74, but not BI-2, binds across monomers within multimerized capsid affecting an inter-hexamer interface that is crucial for maintaining intact virions and that the addition of saturating concentrations of PF74 causes an irreversible block to viral reverse transcription.

Vyšlo v časopise: Host Cofactors and Pharmacologic Ligands Share an Essential Interface in HIV-1 Capsid That Is Lost upon Disassembly. PLoS Pathog 10(10): e32767. doi:10.1371/journal.ppat.1004459
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1004459

Souhrn

Zdroje

1. FrankeEK, YuanHE, LubanJ (1994) Specific incorporation of cyclophilin A into HIV-1 virions. Nature 372 : 359–362.

2. LeeK, AmbroseZ, MartinTD, OztopI, MulkyA, et al. (2010) Flexible use of nuclear import pathways by HIV-1. Cell Host Microbe 7 : 221–233.

3. RasaiyaahJ, TanCP, FletcherAJ, PriceAJ, BlondeauC, et al. (2013) HIV-1 evades innate immune recognition through specific cofactor recruitment. Nature 503 : 402–405.

4. BrassAL, DykxhoornDM, BenitaY, YanN, EngelmanA, et al. (2008) Identification of host proteins required for HIV infection through a functional genomic screen. Science 319 : 921–926.

5. BushmanFD, MalaniN, FernandesJ, D'OrsoI, CagneyG, et al. (2009) Host cell factors in HIV replication: meta-analysis of genome-wide studies. PLoS Pathog 5: e1000437.

6. KonigR, ZhouY, EllederD, DiamondTL, BonamyGM, et al. (2008) Global analysis of host-pathogen interactions that regulate early-stage HIV-1 replication. Cell 135 : 49–60.

7. CherepanovP, MaertensG, ProostP, DevreeseB, Van BeeumenJ, et al. (2003) HIV-1 integrase forms stable tetramers and associates with LEDGF/p75 protein in human cells. The Journal of biological chemistry 278 : 372–381.

8. TowersGJ, HatziioannouT, CowanS, GoffSP, LubanJ, et al. (2003) Cyclophilin A modulates the sensitivity of HIV-1 to host restriction factors. Nat Med 9 : 1138–1143.

9. SchallerT, OcwiejaKE, RasaiyaahJ, PriceAJ, BradyTL, et al. (2011) HIV-1 Capsid-Cyclophilin Interactions Determine Nuclear Import Pathway, Integration Targeting and Replication Efficiency. PLoS Pathog 7: e1002439.

10. KrishnanL, MatreyekKA, OztopI, LeeK, TipperCH, et al. (2010) The requirement for cellular transportin 3 (TNPO3 or TRN-SR2) during infection maps to human immunodeficiency virus type 1 capsid and not integrase. J Virol 84 : 397–406.

11. OcwiejaKE, BradyTL, RonenK, HuegelA, RothSL, et al. (2011) HIV Integration Targeting: A Pathway Involving Transportin-3 and the Nuclear Pore Protein RanBP2. PLoS Pathog 7: e1001313.

12. ChristF, ThysW, De RijckJ, GijsbersR, AlbaneseA, et al. (2008) Transportin-SR2 imports HIV into the nucleus. Curr Biol 18 : 1192–1202.

13. ForsheyBM, von SchwedlerU, SundquistWI, AikenC (2002) Formation of a human immunodeficiency virus type 1 core of optimal stability is crucial for viral replication. J Virol 76 : 5667–5677.

14. HulmeAE, PerezO, HopeTJ (2011) Complementary assays reveal a relationship between HIV-1 uncoating and reverse transcription. Proc Natl Acad Sci U S A 108 : 9975–9980.

15. YamashitaM, EmermanM (2004) Capsid is a dominant determinant of retrovirus infectivity in nondividing cells. J Virol 78 : 5670–5678.

16. BichelK, PriceAJ, SchallerT, TowersGJ, FreundSM, et al. (2013) HIV-1 capsid undergoes coupled binding and isomerization by the nuclear pore protein NUP358. Retrovirology 10 : 81.

17. ZhaoY, ChenY, SchutkowskiM, FischerG, KeH (1997) Cyclophilin A complexed with a fragment of HIV-1 gag protein: insights into HIV-1 infectious activity. Structure 5 : 139–146.

18. BraatenD, LubanJ (2001) Cyclophilin A regulates HIV-1 infectivity, as demonstrated by gene targeting in human T cells. Embo J 20 : 1300–1309.

19. De IacoA, LubanJ (2011) Inhibition of HIV-1 infection by TNPO3 depletion is determined by capsid and detectable after viral cDNA enters the nucleus. Retrovirology 8 : 98.

20. MatreyekKA, YucelSS, LiX, EngelmanA (2013) Nucleoporin NUP153 phenylalanine-glycine motifs engage a common binding pocket within the HIV-1 capsid protein to mediate lentiviral infectivity. PLoS pathogens 9: e1003693.

21. PriceAJ, FletcherAJ, SchallerT, ElliottT, LeeK, et al. (2012) CPSF6 defines a conserved capsid interface that modulates HIV-1 replication. PLoS pathogens 8: e1002896.

22. MatreyekKA, EngelmanA (2011) The requirement for nucleoporin NUP153 during human immunodeficiency virus type 1 infection is determined by the viral capsid. J Virol 85 : 7818–7827.

23. AmbroseZ, AikenC (2014) HIV-1 uncoating: connection to nuclear entry and regulation by host proteins. Virology 454–455C: 371–379.

24. MaertensGN, CookNJ, WangW, HareS, GuptaSS, et al. (2014) Structural basis for nuclear import of splicing factors by human Transportin 3. Proceedings of the National Academy of Sciences of the United States of America 111 : 2728–2733.

25. De IacoA, SantoniF, VannierA, GuipponiM, AntonarakisS, et al. (2013) TNPO3 protects HIV-1 replication from CPSF6-mediated capsid stabilization in the host cell cytoplasm. Retrovirology 10 : 20.

26. YamashitaM, PerezO, HopeTJ, EmermanM (2007) Evidence for direct involvement of the capsid protein in HIV infection of nondividing cells. PLoS Pathog 3 : 1502–1510.

27. BlairWS, PickfordC, IrvingSL, BrownDG, AndersonM, et al. (2010) HIV capsid is a tractable target for small molecule therapeutic intervention. PLoS Pathog 6: e1001220.

28. LamorteL, TitoloS, LemkeCT, GoudreauN, MercierJF, et al. (2013) Discovery of Novel Small-Molecule HIV-1 Replication Inhibitors That Stabilize Capsid Complexes. Antimicrobial agents and chemotherapy 57 : 4622–4631.

29. ShiJ, ZhouJ, ShahVB, AikenC, WhitbyK (2011) Small-molecule inhibition of human immunodeficiency virus type 1 infection by virus capsid destabilization. J Virol 85 : 542–549.

30. HoriT, TakeuchiH, SaitoH, SakumaR, InagakiY, et al. (2013) A carboxy-terminally truncated human CPSF6 lacking residues encoded by exon 6 inhibits HIV-1 cDNA synthesis and promotes capsid disassembly. Journal of virology 87 : 7726–7736.

31. PornillosO, Ganser-PornillosBK, KellyBN, HuaY, WhitbyFG, et al. (2009) X-ray structures of the hexameric building block of the HIV capsid. Cell 137 : 1282–1292.

32. FrickeT, Brandariz-NunezA, WangX, SmithAB3rd, Diaz-GrifferoF (2013) Human cytosolic extracts stabilize the HIV-1 core. Journal of virology 87 : 10587–10597.

33. PornillosO, Ganser-PornillosBK, BanumathiS, HuaY, YeagerM (2010) Disulfide bond stabilization of the hexameric capsomer of human immunodeficiency virus. J Mol Biol 401 : 985–995.

34. ByeonIJ, HouG, HanY, SuiterCL, AhnJ, et al. (2012) Motions on the millisecond time scale and multiple conformations of HIV-1 capsid protein: implications for structural polymorphism of CA assemblies. Journal of the American Chemical Society 134 : 6455–6466.

35. LeeK, MulkyA, YuenW, MartinTD, MeyersonNR, et al. (2012) HIV-1 Capsid-Targeting Domain of Cleavage and Polyadenylation Specificity Factor 6. J Virol 86 : 3851–3860.

36. GallivanJP, DoughertyDA (2000) A Computational Study of Cation-Pi Interactions vs Salt Bridges in Aqueous Media: Implications for Protein Engineering. J Am Chem Soc 122 : 870–874.

37. ShahVB, ShiJ, HoutDR, OztopI, KrishnanL, et al. (2013) The host proteins transportin SR2/TNPO3 and cyclophilin A exert opposing effects on HIV-1 uncoating. Journal of virology 87 : 422–432.

38. HenningMS, DuboseBN, BurseMJ, AikenC, YamashitaM (2014) In vivo functions of CPSF6 for HIV-1 as revealed by HIV-1 capsid evolution in HLA-B27-positive subjects. PLoS pathogens 10: e1003868.

39. KwongPD, DoyleML, CasperDJ, CicalaC, LeavittSA, et al. (2002) HIV-1 evades antibody-mediated neutralization through conformational masking of receptor-binding sites. Nature 420 : 678–682.

40. Collaborative Computational Project N (1994) The CCP4 suite: programs for protein crystallography. Acta Crystallogr D Biol Crystallogr 50 : 760–763.

41. ZennouV, Perez-CaballeroD, GottlingerH, BieniaszPD (2004) APOBEC3G incorporation into human immunodeficiency virus type 1 particles. Journal of virology 78 : 12058–12061.

42. NaldiniL, BlomerU, GallayP, OryD, MulliganR, et al. (1996) In vivo gene delivery and stable transduction of nondividing cells by a lentiviral vector. Science 272 : 263–267.

43. YeeJK, FriedmannT, BurnsJC (1994) Generation of high-titer pseudotyped retroviral vectors with very broad host range. Methods in cell biology 43 Pt A: 99–112.

44. KrissinelE, HenrickK (2007) Inference of macromolecular assemblies from crystalline state. Journal of molecular biology 372 : 774–797.

45. LaskowskiRA, SwindellsMB (2011) LigPlot+: multiple ligand-protein interaction diagrams for drug discovery. Journal of chemical information and modeling 51 : 2778–2786.