Rhesus TRIM5α Disrupts the HIV-1 Capsid at the InterHexamer Interfaces
TRIM proteins play important roles in the innate immune defense against retroviral infection, including human immunodeficiency virus type-1 (HIV-1). Rhesus macaque TRIM5α (TRIM5αrh) targets the HIV-1 capsid and blocks infection at an early post-entry stage, prior to reverse transcription. Studies have shown that binding of TRIM5α to the assembled capsid is essential for restriction and requires the coiled-coil and B30.2/SPRY domains, but the molecular mechanism of restriction is not fully understood. In this study, we investigated, by cryoEM combined with mutagenesis and chemical cross-linking, the direct interactions between HIV-1 capsid protein (CA) assemblies and purified TRIM5αrh containing coiled-coil and SPRY domains (CC-SPRYrh). Concentration-dependent binding of CC-SPRYrh to CA assemblies was observed, while under equivalent conditions the human protein did not bind. Importantly, CC-SPRYrh, but not its human counterpart, disrupted CA tubes in a non-random fashion, releasing fragments of protofilaments consisting of CA hexamers without dissociation into monomers. Furthermore, such structural destruction was prevented by inter-hexamer crosslinking using P207C/T216C mutant CA with disulfide bonds at the CTD-CTD trimer interface of capsid assemblies, but not by intra-hexamer crosslinking via A14C/E45C at the NTD-NTD interface. The same disruption effect by TRIM5αrh on the inter-hexamer interfaces also occurred with purified intact HIV-1 cores. These results provide insights concerning how TRIM5α disrupts the virion core and demonstrate that structural damage of the viral capsid by TRIM5α is likely one of the important components of the mechanism of TRIM5α-mediated HIV-1 restriction.
Vyšlo v časopise:
Rhesus TRIM5α Disrupts the HIV-1 Capsid at the InterHexamer Interfaces. PLoS Pathog 7(3): e32767. doi:10.1371/journal.ppat.1002009
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002009
Souhrn
TRIM proteins play important roles in the innate immune defense against retroviral infection, including human immunodeficiency virus type-1 (HIV-1). Rhesus macaque TRIM5α (TRIM5αrh) targets the HIV-1 capsid and blocks infection at an early post-entry stage, prior to reverse transcription. Studies have shown that binding of TRIM5α to the assembled capsid is essential for restriction and requires the coiled-coil and B30.2/SPRY domains, but the molecular mechanism of restriction is not fully understood. In this study, we investigated, by cryoEM combined with mutagenesis and chemical cross-linking, the direct interactions between HIV-1 capsid protein (CA) assemblies and purified TRIM5αrh containing coiled-coil and SPRY domains (CC-SPRYrh). Concentration-dependent binding of CC-SPRYrh to CA assemblies was observed, while under equivalent conditions the human protein did not bind. Importantly, CC-SPRYrh, but not its human counterpart, disrupted CA tubes in a non-random fashion, releasing fragments of protofilaments consisting of CA hexamers without dissociation into monomers. Furthermore, such structural destruction was prevented by inter-hexamer crosslinking using P207C/T216C mutant CA with disulfide bonds at the CTD-CTD trimer interface of capsid assemblies, but not by intra-hexamer crosslinking via A14C/E45C at the NTD-NTD interface. The same disruption effect by TRIM5αrh on the inter-hexamer interfaces also occurred with purified intact HIV-1 cores. These results provide insights concerning how TRIM5α disrupts the virion core and demonstrate that structural damage of the viral capsid by TRIM5α is likely one of the important components of the mechanism of TRIM5α-mediated HIV-1 restriction.
Zdroje
1. StremlauMOwensCMPerronMJKiesslingMAutissierP 2004 The cytoplasmic body component TRIM5alpha restricts HIV-1 infection in Old World monkeys. Nature 427 848 853
2. YapMWNisoleSLynchCStoyeJP 2004 Trim5alpha protein restricts both HIV-1 and murine leukemia virus. Proc Natl Acad Sci U S A 101 10786 10791
3. StremlauMPerronMLeeMLiYSongB 2006 Specific recognition and accelerated uncoating of retroviral capsids by the TRIM5alpha restriction factor. Proc Natl Acad Sci U S A 103 5514 5519
4. PerronMJStremlauMLeeMJavanbakhtHSongB 2007 The human TRIM5alpha restriction factor mediates accelerated uncoating of the N-tropic murine leukemia virus capsid. J Virol 81 2138 2148
5. ShiJAikenC 2006 Saturation of TRIM5 alpha-mediated restriction of HIV-1 infection depends on the stability of the incoming viral capsid. Virology 350 493 500
6. SebastianSLubanJ 2005 TRIM5alpha selectively binds a restriction-sensitive retroviral capsid. Retrovirology 2 40
7. ReymondAMeroniGFantozziAMerlaGCairoS 2001 The tripartite motif family identifies cell compartments. EMBO J 20 2140 2151
8. NisoleSStoyeJPSaibA 2005 Trim family proteins: Retroviral restriction and antiviral defence. Nat Rev Microbiol 3 799 808
9. OzatoKShinDMChangTHMorseHC3rd 2008 TRIM family proteins and their emerging roles in innate immunity. Nat Rev Immunol 8 849 860
10. StremlauMPerronMWelikalaSSodroskiJ 2005 Species-specific variation in the B30.2(SPRY) domain of TRIM5 alpha determines the potency of human immunodeficiency virus restriction. J Virol 79 3139 3145
11. YapMWNisoleSStoyeJP 2005 A single amino acid change in the SPRY domain of human Trim5 alpha leads to HIV-1 restriction. Curr Biol 15 73 78
12. Perez-CaballeroDHatziioannouTYangACowanSBieniaszPD 2005 Human tripartite motif 5 alpha domains responsible for retrovirus restriction activity and specificity. J Virol 79 8969 8978
13. SawyerSLWuLIEmermanMMalikHS 2005 Positive selection of primate TRIM5alpha identifies a critical species-specific retroviral restriction domain. Proc Natl Acad Sci U S A 102 2832 2837
14. OhkuraSYapMWSheldonTStoyeJP 2006 All three variable regions of the TRIM5alpha B30.2 domain can contribute to the specificity of retrovirus restriction. J Virol 80 8554 8565
15. SongBGoldBO'HuiginCJavanbakhtHLiX 2005 The B30.2(SPRY) domain of the retroviral restriction factor TRIM5alpha exhibits lineage-specific length and sequence variation in primates. J Virol 79 6111 6121
16. JamesLCKeebleAHKhanZRhodesDATrowsdaleJ 2007 Structural basis for PRYSPRY-mediated tripartite motif (TRIM) protein function. Proc Natl Acad Sci U S A 104 6200 6205
17. LiYLiXStremlauMLeeMSodroskiJ 2006 Removal of arginine 332 allows human TRIM5alpha to bind human immunodeficiency virus capsids and to restrict infection. J Virol 80 6738 6744
18. MischeCCJavanbakhtHSongBDiaz-GrifferoFStremlauM 2005 Retroviral restriction factor TRIM5alpha is a trimer. J Virol 79 14446 14450
19. JavanbakhtHYuanWYeungDFSongBDiaz-GrifferoF 2006 Characterization of TRIM5alpha trimerization and its contribution to human immunodeficiency virus capsid binding. Virology 353 234 246
20. MaillardPVEccoGOrtizMTronoD 2010 The specificity of TRIM5alpha-mediated restriction is influenced by its coiled-coil domain. J Virol 84 5790 801
21. LiXSodroskiJ 2008 The TRIM5alpha B-box 2 Domain Promotes Cooperative Binding to the Retroviral Capsid by Mediating Higher-order Self-association. J Virol 82 11495 502
22. Diaz-GrifferoFQinXRHayashiFKigawaTFinziA 2009 A B-Box 2 Surface Patch Important for TRIM5 alpha Self-Association, Capsid Binding Avidity, and Retrovirus Restriction. J Virol 83 10737 10751
23. JavanbakhtHDiaz-GrifferoFStremlauMSiZSodroskiJ 2005 The contribution of RING and B-box 2 domains to retroviral restriction mediated by monkey TRIM5alpha. J Biol Chem 280 26933 26940
24. Diaz-GrifferoFKarAPerronMXiangSHJavanbakhtH 2007 Modulation of retroviral restriction and proteasome inhibitor-resistant turnover by changes in the TRIM5alpha B-box 2 domain. J Virol 81 10362 10378
25. MeroniGDiez-RouxG 2005 TRIM/RBCC, a novel class of ‘single protein RING finger’ E3 ubiquitin ligases. Bioessays 27 1147 1157
26. Perez-CaballeroDHatziioannouTZhangFCowanSBieniaszPD 2005 Restriction of human immunodeficiency virus type 1 by TRIM-CypA occurs with rapid kinetics and independently of cytoplasmic bodies, ubiquitin, and proteasome activity. J Virol 79 15567 15572
27. WuXAndersonJLCampbellEMJosephAMHopeTJ 2006 Proteasome inhibitors uncouple rhesus TRIM5alpha restriction of HIV-1 reverse transcription and infection. Proc Natl Acad Sci U S A 103 7465 7470
28. RoldCJAikenC 2008 Proteasomal degradation of TRIM5alpha during retrovirus restriction. PLoS Pathog 4 e1000074
29. BerthouxLSebastianSSayahDMLubanJ 2005 Disruption of human TRIM5alpha antiviral activity by nonhuman primate orthologues. J Virol 79 7883 7888
30. MunkCBrandtSMLuceroGLandauNR 2002 A dominant block to HIV-1 replication at reverse transcription in simian cells. Proc Natl Acad Sci U S A 99 13843 13848
31. DoddingMPBockMYapMWStoyeJP 2005 Capsid processing requirements for abrogation of Fv1 and Ref1 restriction. J Virol 79 10571 10577
32. LangelierCRSandrinVEckertDMChristensenDEChandrasekaranV 2008 Biochemical characterization of a recombinant TRIM5alpha protein that restricts human immunodeficiency virus type 1 replication. J Virol 82 11682 11694
33. KarAKDiaz-GrifferoFLiYLiXSodroskiJ 2008 Biochemical and Biophysical Characterization of a Chimeric TRIM21-TRIM5alpha Protein. J Virol 82 11669 81
34. Ganser-PornillosBKChandrasekaranVPornillosOSodroskiJGSundquistWI 2011 Hexagonal assembly of a restricting TRIM5alpha protein. Proc Natl Acad Sci U S A.108 534 539
35. Diaz-GrifferoFLiXJavanbakhtHSongBWelikalaS 2006 Rapid turnover and polyubiquitylation of the retroviral restriction factor TRIM5. Virology 349 300 315
36. LiXLiYStremlauMYuanWSongB 2006 Functional replacement of the RING, B-box 2, and coiled-coil domains of tripartite motif 5alpha (TRIM5alpha) by heterologous TRIM domains. J Virol 80 6198 6206
37. BlackLRAikenC 2010 TRIM5alpha disrupts the structure of assembled HIV-1 capsid complexes in vitro. J Virol 84 6564 6569
38. ByeonIJMengXJungJZhaoGYangR 2009 Structural convergence between Cryo-EM and NMR reveals intersubunit interactions critical for HIV-1 capsid function. Cell 139 780 790
39. OwensCMSongBPerronMJYangPCStremlauM 2004 Binding and susceptibility to postentry restriction factors in monkey cells are specified by distinct regions of the human immunodeficiency virus type 1 capsid. J Virol 78 5423 5437
40. PornillosOGanser-PornillosBKKellyBNHuaYWhitbyFG 2009 X-ray structures of the hexameric building block of the HIV capsid. Cell 137 1282 1292
41. Ganser-PornillosBKChengAYeagerM 2007 Structure of full-length HIV-1 CA: a model for the mature capsid lattice. Cell 131 70 79
42. DesaiAMitchisonTJ 1997 Microtubule polymerization dynamics. Annu Rev Cell Dev Biol 13 83 117
43. NogalesEWangHW 2006 Structural intermediates in microtubule assembly and disassembly: how and why? Curr Opin Cell Biol 18 179 184
44. FujiwaraITakahashiSTadakumaHFunatsuTIshiwataS 2002 Microscopic analysis of polymerization dynamics with individual actin filaments. Nat Cell Biol 4 666 673
45. HinshawJESchmidSL 1995 Dynamin self-assembles into rings suggesting a mechanism for coated vesicle budding. Nature 374 190 192
46. ForsheyBMvon SchwedlerUSundquistWIAikenC 2002 Formation of a human immunodeficiency virus type 1 core of optimal stability is crucial for viral replication. J Virol 76 5667 5677
47. Diaz-GrifferoFKarALeeMStremlauMPoeschlaE 2007 Comparative requirements for the restriction of retrovirus infection by TRIM5alpha and TRIMCyp. Virology 369 400 410
48. YapMWMortuzaGBTaylorIAStoyeJP 2007 The design of artificial retroviral restriction factors. Virology 365 302 314
49. GanserBKLiSKlishkoVYFinchJTSundquistWI 1999 Assembly and analysis of conical models for the HIV-1 core. Science 283 80 83
50. AikenC 2009 Cell-free assays for HIV-1 uncoating. Methods Mol Biol 485 41 53
51. GallayPHopeTChinDTronoD 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
52. DurocherYPerretSKamenA 2002 High-level and high-throughput recombinant protein production by transient transfection of suspension-growing human 293-EBNA1 cells. Nucleic Acids Res 30 E9
53. WehrlyKChesebroB 1997 p24 antigen capture assay for quantification of human immunodeficiency virus using readily available inexpensive reagents. Methods 12 288 293
54. PhillipsJMMurrayPSMurrayDVogtVM 2008 A molecular switch required for retrovirus assembly participates in the hexagonal immature lattice. EMBO J 27 1411 1420
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2011 Číslo 3
- Parazitičtí červi v terapii Crohnovy choroby a dalších zánětlivých autoimunitních onemocnění
- Koronavirus hýbe světem: Víte jak se chránit a jak postupovat v případě podezření?
- Očkování proti virové hemoragické horečce Ebola experimentální vakcínou rVSVDG-ZEBOV-GP
Najčítanejšie v tomto čísle
- A Toxin that Hijacks the Host Ubiquitin Proteolytic System
- Blood Meal-Derived Heme Decreases ROS Levels in the Midgut of and Allows Proliferation of Intestinal Microbiota
- HIV Integration Targeting: A Pathway Involving Transportin-3 and the Nuclear Pore Protein RanBP2
- Invasive Extravillous Trophoblasts Restrict Intracellular Growth and Spread of