A Cyclophilin Homology Domain-Independent Role for Nup358 in HIV-1 Infection

English version České info

The large nucleoporin Nup358/RanBP2 forms eight filaments that project from the nuclear pore into the cytoplasm where they function as docking platforms for nucleocytoplasmic transport receptors. RNAi screens have implicated Nup358 in the HIV-1 life cycle. The 164 C-terminal amino acids of this 3,224 amino acid protein are a cyclophilin homology domain (Nup358Cyp), which has potential to bind the HIV-1 capsid and regulate viral progress to integration. Here we examined the virological role of Nup358 in conditional knockout mouse cells and in RNAi-depleted human CD4+ T cells. Cre-mediated gene knockout was toxic and diminished HIV-1 infectivity. However, cellular health and HIV-1 susceptibility were coordinately preserved if, prior to gene inactivation, a transposon was used to express all of Nup358 or only the N-terminal 1340 amino acids that contain three FG repeats and a Ran-binding domain. HIV-1, but not N74D capsid-mutant HIV-1, was markedly sensitive to TNPO3 depletion, but they infected 1–1340 segment-complemented Nup358 knockout cells equivalently. Human and mouse CypA both rescued HIV-1 in CypA gene −/− Jurkat cells and TRIM-Nup358Cyp fusions derived from each species were equally antiviral; each also inhibited both WT and N74D virus. In the human CD4+ T cell line SupT1, abrupt Nup358 depletion reduced viral replication but stable Nup358-depleted cells replicated HIV-1 normally. Thus, human CD4+ T cells can accommodate to loss of Nup358 and preserve HIV-1 susceptibility. Experiments with cylosporine, viruses with capsids that do not bind cyclophilins, and growth arrest did not uncover viral dependency on the C-terminal domains of Nup358. Our data reinforce the virological importance of TNPO3 and show that Nup358 supports nuclear transport functions important for cellular homeostasis and for HIV-1 nuclear import. However, the results do not suggest direct roles for the Nup358 cyclophilin or SUMO E3 ligase domains in engaging the HIV-1 capsid prior to nuclear translocation.

Vyšlo v časopise: A Cyclophilin Homology Domain-Independent Role for Nup358 in HIV-1 Infection. PLoS Pathog 10(2): e32767. doi:10.1371/journal.ppat.1003969
Kategorie: Research Article
prolekare.web.journal.doi_sk: https://doi.org/10.1371/journal.ppat.1003969

Souhrn

Zdroje

1. NarayanO, WolinskyJS, ClementsJE, StrandbergJD, GriffinDE, et al. (1982) Slow virus replication: the role of macrophages in the persistence and expression of visna viruses of sheep and goats. J Gen Virol 59: 345–356.

2. WeinbergJB, MatthewsTJ, CullenBR, MalimMH (1991) Productive human immunodeficiency virus type 1 (HIV-1) infection of nonproliferating human monocytes. Journal of Experimental Medicine 174: 1477–1482.

3. GoldstoneDC, YapMW, RobertsonLE, HaireLF, TaylorWR, et al. (2010) Structural and functional analysis of prehistoric lentiviruses uncovers an ancient molecular interface. Cell Host Microbe 8: 248–259.

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

5. PoeschlaE, Wong-StaalF, LooneyD (1998) Efficient transduction of nondividing cells by feline immunodeficiency virus lentiviral vectors. Nature Medicine 4: 354–357.

6. SuzukiY, CraigieR (2007) The road to chromatin - nuclear entry of retroviruses. Nat Rev Microbiol 5: 187–196.

7. FouchierRA, MeyerBE, SimonJH, FischerU, MalimMH (1997) HIV-1 infection of non-dividing cells: evidence that the amino-terminal basic region of the viral matrix protein is important for Gag processing but not for post-entry nuclear import. Embo J 16: 4531–4539.

8. ReilH, BukovskyAA, GelderblomHR, GottlingerHG (1998) Efficient HIV-1 replication can occur in the absence of the viral matrix protein. Embo J 17: 2699–2708.

9. YamashitaM, EmermanM (2005) The cell cycle independence of HIV infections is not determined by known karyophilic viral elements. PLoS Pathog 1: e18.

10. DvorinJD, BellP, MaulGG, YamashitaM, EmermanM, et al. (2002) Reassessment of the roles of integrase and the central DNA flap in human immunodeficiency virus type 1 nuclear import. J Virol 76: 12087–12096.

11. LimonA, NakajimaN, LuR, GhoryHZ, EngelmanA (2002) Wild-type levels of nuclear localization and human immunodeficiency virus type 1 replication in the absence of the central DNA flap. J Virol 76: 12078–12086.

12. HuC, SaenzDT, FadelHJ, WalkerW, PeretzM, et al. (2010) The HIV-1 central polypurine tract functions as a second line of defense against APOBEC3G/F. J Virol 84: 11981–11993.

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

14. DismukeDJ, AikenC (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.

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

16. AlberF, DokudovskayaS, VeenhoffLM, ZhangW, KipperJ, et al. (2007) Determining the architectures of macromolecular assemblies. Nature 450: 683–694.

17. TerryLJ, WenteSR (2009) Flexible gates: dynamic topologies and functions for FG nucleoporins in nucleocytoplasmic transport. Eukaryot Cell 8: 1814–1827.

18. ChookYM, SuelKE (2010) Nuclear import by karyopherin-betas: recognition and inhibition. Biochim Biophys Acta 1813: 1593–1606.

19. BrassAL, DykxhoornDM, BenitaY, YanN, EngelmanA, et al. (2008) Identification of Host Proteins Required for HIV Infection Through a Functional Genomic Screen. Science 319 (5865) 921–6.

20. 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.

21. FassatiA, GorlichD, HarrisonI, ZaytsevaL, MingotJM (2003) Nuclear import of HIV-1 intracellular reverse transcription complexes is mediated by importin 7. Embo J 22: 3675–3685.

22. KrishnanL, MatreyekKA, OztopI, LeeK, TipperCH, et al. (2009) 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.

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

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

25. 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.

26. ZhangR, MehlaR, ChauhanA (2010) Perturbation of host nuclear membrane component RanBP2 impairs the nuclear import of human immunodeficiency virus -1 preintegration complex (DNA). PLoS One 5: e15620.

27. ZhouL, SokolskajaE, JollyC, JamesW, CowleySA, et al. (2011) Transportin 3 promotes a nuclear maturation step required for efficient HIV-1 integration. PLoS Pathog 7: e1002194.

28. 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.

29. 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.

30. WuJ, MatunisMJ, KraemerD, BlobelG, CoutavasE (1995) Nup358, a cytoplasmically exposed nucleoporin with peptide repeats, Ran-GTP binding sites, zinc fingers, a cyclophilin A homologous domain, and a leucine-rich region. J Biol Chem 270: 14209–14213.

31. XylourgidisN, FornerodM (2009) Acting out of character: regulatory roles of nuclear pore complex proteins. Dev Cell 17: 617–625.

32. WangP, HeitmanJ (2005) The cyclophilins. Genome Biol 6: 226.

33. LubanJ, BossoltKL, FrankeEK, KalpanaGV, GoffSP (1993) Human immunodeficiency virus type 1 Gag protein binds to cyclophilins A and B. Cell 73: 1067–1078.

34. BraatenD, AberhamC, FrankeEK, YinL, PharesW, et al. (1996) Cyclosporine A-resistant human immunodeficiency virus type 1 mutants demonstrate that Gag encodes the functional target of cyclophilin A. J Virol 70: 5170–5176.

35. BraatenD, FrankeEK, LubanJ (1996) Cyclophilin A is required for an early step in the life cycle of human immunodeficiency virus type 1 before the initiation of reverse transcription. J Virol 70: 3551–3560.

36. ThaliM, BukovskyA, KondoE, RosenwirthB, WalshCT, et al. (1994) Functional association of cyclophilin A with HIV-1 virions. Nature 372: 363–365.

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

38. KootstraNA, MunkC, TonnuN, LandauNR, VermaIM (2003) Abrogation of postentry restriction of HIV-1-based lentiviral vector transduction in simian cells. Proc Natl Acad Sci U S A 100: 1298–1303.

39. 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.

40. SokolskajaE, LubanJ (2006) Cyclophilin, TRIM5, and innate immunity to HIV-1. Curr Opin Microbiol 9: 404–408.

41. TowersGJ (2007) The control of viral infection by tripartite motif proteins and cyclophilin A. Retrovirology 4: 40.

42. BerthouxL, SebastianS, SokolskajaE, LubanJ (2005) Cyclophilin A is required for TRIM5{alpha}-mediated resistance to HIV-1 in Old World monkey cells. Proc Natl Acad Sci U S A 102: 14849–14853.

43. KeckesovaZ, YlinenLM, TowersGJ (2006) Cyclophilin A renders human immunodeficiency virus type 1 sensitive to Old World monkey but not human TRIM5 alpha antiviral activity. J Virol 80: 4683–4690.

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

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

46. LahayeX, SatohT, GentiliM, CerboniS, ConradC, et al. (2013) The Capsids of HIV-1 and HIV-2 Determine Immune Detection of the Viral cDNA by the Innate Sensor cGAS in Dendritic Cells. Immunity 39: 1132–1142.

47. PichlerA, GastA, SeelerJS, DejeanA, MelchiorF (2002) The nucleoporin RanBP2 has SUMO1 E3 ligase activity. Cell 108: 109–120.

48. DawlatyMM, MalureanuL, JeganathanKB, KaoE, SustmannC, et al. (2008) Resolution of sister centromeres requires RanBP2-mediated SUMOylation of topoisomerase IIalpha. Cell 133: 103–115.

49. GurerC, BerthouxL, LubanJ (2005) Covalent modification of human immunodeficiency virus type 1 p6 by SUMO-1. J Virol 79: 910–917.

50. ZamborliniA, CoifficA, BeauclairG, DelelisO, ParisJ, et al. (2011) Impairment of human immunodeficiency virus type-1 integrase SUMOylation correlates with an early replication defect. J Biol Chem 286: 21013–21022.

51. JosephJ, DassoM (2008) The nucleoporin Nup358 associates with and regulates interphase microtubules. FEBS Lett 582: 190–196.

52. McDonaldD, VodickaMA, LuceroG, SvitkinaTM, BorisyGG, et al. (2002) Visualization of the intracellular behavior of HIV in living cells. J Cell Biol 159: 441–452.

53. SayahDM, SokolskajaE, BerthouxL, LubanJ (2004) Cyclophilin A retrotransposition into TRIM5 explains owl monkey resistance to HIV-1. Nature 430: 569–573.

54. 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.

55. LinDH, ZimmermannS, StuweT, StuweE, HoelzA (2013) Structural and Functional Analysis of the C-Terminal Domain of Nup358/RanBP2. J Mol Biol 425: 1318–1329.

56. LandryJ, PylP, RauschT, ZichnerT, TekkedilM, et al. (2013) The genomic and transcriptomic landscape of a HeLa cell line. G3: Genes, Genomes and Genetics 3: 1213–1224.

57. AslanukovA, BhowmickR, GurujuM, OswaldJ, RazD, et al. (2006) RanBP2 modulates Cox11 and hexokinase I activities and haploinsufficiency of RanBP2 causes deficits in glucose metabolism. PLoS Genet 2: e177.

58. HamadaM, HaegerA, JeganathanKB, van ReeJH, MalureanuL, et al. (2011) Ran-dependent docking of importin-beta to RanBP2/Nup358 filaments is essential for protein import and cell viability. J Cell Biol 194: 597–612.

59. RibbeckK, GorlichD (2001) Kinetic analysis of translocation through nuclear pore complexes. The EMBO journal 20: 1320–1330.

60. WaldeS, ThakarK, HuttenS, SpillnerC, NathA, et al. (2011) The Nucleoporin Nup358/RanBP2 Promotes Nuclear Import in a Cargo- and Transport Receptor-Specific Manner. Traffic 13: 218–233.

61. BalciunasD, WangensteenKJ, WilberA, BellJ, GeurtsA, et al. (2006) Harnessing a high cargo-capacity transposon for genetic applications in vertebrates. PLoS Genet 2: e169.

62. ThysW, De HouwerS, DemeulemeesterJ, TaltynovO, VancraenenbroeckR, et al. (2011) Interplay between HIV entry and transportin-SR2 dependency. Retrovirology 8: 7.

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

64. 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.

65. LarueR, GuptaK, WuenschC, ShkriabaiN, KesslJJ, et al. (2012) Interaction of the HIV-1 intasome with transportin 3 protein (TNPO3 or TRN-SR2). J Biol Chem 287: 34044–34058.

66. Valle-CasusoJC, Di NunzioF, YangY, ReszkaN, LienlafM, et al. (2012) TNPO3 is required for HIV-1 replication after nuclear import but prior to integration and binds the HIV-1 core. J Virol 86: 5931–5936.

67. 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.

68. FrickeT, Valle-CasusoJC, WhiteTE, Brandariz-NunezA, BoscheWJ, et al. (2013) The ability of TNPO3-depleted cells to inhibit HIV-1 infection requires CPSF6. Retrovirology 10: 46.

69. LubanJ (2007) Cyclophilin A, TRIM5, and resistance to human immunodeficiency virus type 1 infection. J Virol 81: 1054–1061.

70. van ValenL (1991) HeLa, a new microbial species. Evolutionary Theory and Review 10: 71–74.

71. 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 Pathog 9: e1003693.

72. OriA, BanterleN, IskarM, Andres-PonsA, EscherC, et al. (2013) Cell type-specific nuclear pores: a case in point for context-dependent stoichiometry of molecular machines. Mod Pathol 9: 648.

73. LevinA, HayoukaZ, FriedlerA, LoyterA (2011) Transportin 3 and importin alpha are required for effective nuclear import of HIV-1 integrase in virus-infected cells. Nucleus 1: 422–431.

74. GallayP, HopeT, ChinD, TronoD (1997) HIV-1 infection of nondividing cells through the recognition of integrase by the importin/karyopherin pathway. Proc Natl Acad Sci U S A 94: 9825–9830.

75. HuttenS, WaldeS, SpillnerC, HauberJ, KehlenbachRH (2009) The nuclear pore component Nup358 promotes transportin-dependent nuclear import. J Cell Sci 122: 1100–1110.

76. SaitohN, SakamotoC, HagiwaraM, Agredano-MorenoLT, Jimenez-GarciaLF, et al. (2012) The distribution of phosphorylated SR proteins and alternative splicing are regulated by RANBP2. Mol Biol Cell 23: 1115–1128.

77. Di NunzioF, DanckaertA, FrickeT, PerezP, FernandezJ, et al. (2012) Human nucleoporins promote HIV-1 docking at the nuclear pore, nuclear import and integration. PLoS One 7: e46037.

78. AmbroseZ, LeeK, NdjomouJ, XuH, OztopI, et al. (2012) Human immunodeficiency virus type 1 capsid mutation N74D alters cyclophilin A dependence and impairs macrophage infection. J Virol 86: 4708–4714.

79. ShunMC, RaghavendraNK, VandegraaffN, DaigleJE, HughesS, et al. (2007) LEDGF/p75 functions downstream from preintegration complex formation to effect gene-specific HIV-1 integration. Genes Dev 21: 1767–1778.

80. LlanoM, SaenzDT, MeehanA, WongthidaP, PeretzM, et al. (2006) An Essential Role for LEDGF/p75 in HIV Integration. Science 314: 461–464.

81. Llano M, Gaznick N, Poeschla EM (2008) Rapid, controlled and intensive lentiviral vector-based RNAi. In: Kalpana G, Prasad V, editors. HIV Protocols, second edition. Totowa, NJ: Humana Press.

82. SaenzDT, BarrazaR, LoewenN, TeoW, PoeschlaEM (2012) Production, harvest, and concentration of feline immunodeficiency virus-based lentiviral vector from cells grown in CF10 or CF2 devices. Cold Spring Harb Protoc 2012: 118–123.

83. MeehanAM, SaenzDT, MorrisonJH, Garcia-RiveraJA, PeretzM, et al. (2009) LEDGF/p75 proteins with alternative chromatin tethers are functional HIV-1 cofactors. PLoS Pathogens 5: e1000522.