A Host Small GTP-binding Protein ARL8 Plays Crucial Roles in Tobamovirus RNA Replication
Tomato mosaic virus (ToMV), like other eukaryotic positive-strand RNA viruses, replicates its genomic RNA in replication complexes formed on intracellular membranes. Previous studies showed that a host seven-pass transmembrane protein TOM1 is necessary for efficient ToMV multiplication. Here, we show that a small GTP-binding protein ARL8, along with TOM1, is co-purified with a FLAG epitope-tagged ToMV 180K replication protein from solubilized membranes of ToMV-infected tobacco (Nicotiana tabacum) cells. When solubilized membranes of ToMV-infected tobacco cells that expressed FLAG-tagged ARL8 were subjected to immunopurification with anti-FLAG antibody, ToMV 130K and 180K replication proteins and TOM1 were co-purified and the purified fraction showed RNA-dependent RNA polymerase activity that transcribed ToMV RNA. From uninfected cells, TOM1 co-purified with FLAG-tagged ARL8 less efficiently, suggesting that a complex containing ToMV replication proteins, TOM1, and ARL8 are formed on membranes in infected cells. In Arabidopsis thaliana, ARL8 consists of four family members. Simultaneous mutations in two specific ARL8 genes completely inhibited tobamovirus multiplication. In an in vitro ToMV RNA translation-replication system, the lack of either TOM1 or ARL8 proteins inhibited the production of replicative-form RNA, indicating that TOM1 and ARL8 are required for efficient negative-strand RNA synthesis. When ToMV 130K protein was co-expressed with TOM1 and ARL8 in yeast, RNA 5′-capping activity was detected in the membrane fraction. This activity was undetectable or very weak when the 130K protein was expressed alone or with either TOM1 or ARL8. Taken together, these results suggest that TOM1 and ARL8 are components of ToMV RNA replication complexes and play crucial roles in a process toward activation of the replication proteins' RNA synthesizing and capping functions.
Vyšlo v časopise:
A Host Small GTP-binding Protein ARL8 Plays Crucial Roles in Tobamovirus RNA Replication. PLoS Pathog 7(12): e32767. doi:10.1371/journal.ppat.1002409
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002409
Souhrn
Tomato mosaic virus (ToMV), like other eukaryotic positive-strand RNA viruses, replicates its genomic RNA in replication complexes formed on intracellular membranes. Previous studies showed that a host seven-pass transmembrane protein TOM1 is necessary for efficient ToMV multiplication. Here, we show that a small GTP-binding protein ARL8, along with TOM1, is co-purified with a FLAG epitope-tagged ToMV 180K replication protein from solubilized membranes of ToMV-infected tobacco (Nicotiana tabacum) cells. When solubilized membranes of ToMV-infected tobacco cells that expressed FLAG-tagged ARL8 were subjected to immunopurification with anti-FLAG antibody, ToMV 130K and 180K replication proteins and TOM1 were co-purified and the purified fraction showed RNA-dependent RNA polymerase activity that transcribed ToMV RNA. From uninfected cells, TOM1 co-purified with FLAG-tagged ARL8 less efficiently, suggesting that a complex containing ToMV replication proteins, TOM1, and ARL8 are formed on membranes in infected cells. In Arabidopsis thaliana, ARL8 consists of four family members. Simultaneous mutations in two specific ARL8 genes completely inhibited tobamovirus multiplication. In an in vitro ToMV RNA translation-replication system, the lack of either TOM1 or ARL8 proteins inhibited the production of replicative-form RNA, indicating that TOM1 and ARL8 are required for efficient negative-strand RNA synthesis. When ToMV 130K protein was co-expressed with TOM1 and ARL8 in yeast, RNA 5′-capping activity was detected in the membrane fraction. This activity was undetectable or very weak when the 130K protein was expressed alone or with either TOM1 or ARL8. Taken together, these results suggest that TOM1 and ARL8 are components of ToMV RNA replication complexes and play crucial roles in a process toward activation of the replication proteins' RNA synthesizing and capping functions.
Zdroje
1. SchwartzMChenJJandaMSullivanMden BoonJ 2002 A positive-strand RNA virus replication complex parallels form and function of retrovirus capsids. Mol Cell 9 505 514
2. KushnerDBLindenbachBDGrdzelishviliVZNoueiryAOPaulSM 2003 Systematic, genome-wide identification of host genes affecting replication of a positive-strand RNA virus. Proc Natl Acad Sci U S A 100 15764 15769
3. PanavasTServieneEBrasherJNagyPD 2005 Yeast genome-wide screen reveals dissimilar sets of host genes affecting replication of RNA viruses. Proc Natl Acad Sci U S A 102 7326 7331
4. KrishnanMNNgASukumaranBGilfoyFDUchilPD 2008 RNA interference screen for human genes associated with West Nile virus infection. Nature 455 242 245
5. SessionsOMBarrowsNJSouza-NetoJARobinsonTJHersheyCL 2009 Discovery of insect and human dengue virus host factors. Nature 458 1047 1050
6. TaiAWBenitaYPengLFKimSSSakamotoN 2009 A functional genomic screen identifies cellular cofactors of hepatitis C virus replication. Cell Host Microbe 5 298 307
7. PoganyJStorkJLiZNagyPD 2008 In vitro assembly of the Tomato bushy stunt virus replicase requires the host heat shock protein 70. Proc Natl Acad Sci U S A 105 19956 19961
8. TomitaYMizunoTDiezJNaitoSAhlquistP 2003 Mutation of host DnaJ homolog inhibits brome mosaic virus negative-strand RNA synthesis. J Virol 77 2990 2997
9. WeeksSAMillerDJ 2008 The heat shock protein 70 cochaperone YDJ1 is required for efficient membrane-specific flock house virus RNA replication complex assembly and function in Saccharomyces cerevisiae. J Virol 82 2004 2012
10. CastorenaKMWeeksSAStaplefordKACadwalladerAMMillerDJ 2007 A functional heat shock protein 90 chaperone is essential for efficient flock house virus RNA polymerase synthesis in Drosophila cells. J Virol 81 8412 8420
11. WatashiKIshiiNHijikataMInoueDMurataT 2005 Cyclophilin B is a functional regulator of hepatitis C virus RNA polymerase. Mol Cell 19 111 122
12. LiZPoganyJTupmanSEspositoAMKinzyTG 2010 Translation elongation factor 1A facilitates the assembly of the tombusvirus replicase and stimulates minus-strand synthesis. PLoS Pathog 6 e1001175
13. WangRYNagyPD 2008 Tomato bushy stunt virus co-opts the RNA-binding function of a host metabolic enzyme for viral genomic RNA synthesis. Cell Host Microbe 3 178 187
14. DiazAWangXAhlquistP 2010 Membrane-shaping host reticulon proteins play crucial roles in viral RNA replication compartment formation and function. Proc Natl Acad Sci U S A 107 16291 16296
15. BarajasDJiangYNagyPD 2009 A unique role for the host ESCRT proteins in replication of Tomato bushy stunt virus. PLoS Pathog 5 e1000705
16. LeeWMIshikawaMAhlquistP 2001 Mutation of host delta9 fatty acid desaturase inhibits brome mosaic virus RNA replication between template recognition and RNA synthesis. J Virol 75 2097 2106
17. SharmaMSasvariZNagyPD 2010 Inhibition of sterol biosynthesis reduces tombusvirus replication in yeast and plants. J Virol 84 2270 2281
18. MackenzieJMKhromykhAAPartonRG 2007 Cholesterol manipulation by West Nile virus perturbs the cellular immune response. Cell Host Microbe 2 229 239
19. AholaTLampioAAuvinenPKaariainenL 1999 Semliki Forest virus mRNA capping enzyme requires association with anionic membrane phospholipids for activity. EMBO J 18 3164 3172
20. HsuNYIlnytskaOBelovGSantianaMChenYH 2010 Viral reorganization of the secretory pathway generates distinct organelles for RNA replication. Cell 141 799 811
21. ReissSRebhanIBackesPRomero-BreyIErfleH 2011 Recruitment and activation of a lipid kinase by hepatitis C virus NS5A is essential for integrity of the membranous replication compartment. Cell Host Microbe 9 32 45
22. LeeWMAhlquistP 2003 Membrane synthesis, specific lipid requirements, and localized lipid composition changes associated with a positive-strand RNA virus RNA replication protein. J Virol 77 12819 12828
23. HeatonNSPereraRBergerKLKhadkaSLacountDJ 2010 Dengue virus nonstructural protein 3 redistributes fatty acid synthase to sites of viral replication and increases cellular fatty acid synthesis. Proc Natl Acad Sci U S A 107 17345 17350
24. IshikawaMOkadaY 2004 Replication of tobamovirus RNA. Proc Jpn Acad Ser B 80 215 224
25. IshibashiKNishikioriMIshikawaM 2010 Interactions between tobamovirus replication proteins and cellular factors: their impacts on virus multiplication. Mol Plant Microbe Interact 23 1413 1419
26. KomodaKMawatariNHagiwara-KomodaYNaitoSIshikawaM 2007 Identification of a ribonucleoprotein intermediate of tomato mosaic virus RNA replication complex formation. J Virol 81 2584 2591
27. NishikioriMDohiKMoriMMeshiTNaitoS 2006 Membrane-bound tomato mosaic virus replication proteins participate in RNA synthesis and are associated with host proteins in a pattern distinct from those that are not membrane bound. J Virol 80 8459 8468
28. Hagiwara-KomodaYHiraiKMochizukiANishiguchiMMeshiT 2008 Overexpression of a host factor TOM1 inhibits tomato mosaic virus propagation and suppression of RNA silencing. Virology 376 132 139
29. KuriharaYInabaNKutsunaNTakedaATagamiY 2007 Binding of tobamovirus replication protein with small RNA duplexes. J Gen Virol 88 2347 2352
30. AliyariRDingSW 2009 RNA-based viral immunity initiated by the Dicer family of host immune receptors. Immunol Rev 227 176 188
31. YamajiYSakuraiKHamadaKKomatsuKOzekiJ 2010 Significance of eukaryotic translation elongation factor 1A in tobacco mosaic virus infection. Arch Virol 155 263 268
32. YamajiYKobayashiTHamadaKSakuraiKYoshiiA 2006 In vivo interaction between Tobacco mosaic virus RNA-dependent RNA polymerase and host translation elongation factor 1A. Virology 347 100 108
33. TsujimotoYNumagaTOhshimaKYanoMAOhsawaR 2003 Arabidopsis TOBAMOVIRUS MULTIPLICATION (TOM) 2 locus encodes a transmembrane protein that interacts with TOM1. EMBO J 22 335 343
34. YamanakaTOhtaTTakahashiMMeshiTSchmidtR 2000 TOM1, an Arabidopsis gene required for efficient multiplication of a tobamovirus, encodes a putative transmembrane protein. Proc Natl Acad Sci U S A 97 10107 10112
35. OsmanTABuckKW 1997 The tobacco mosaic virus RNA polymerase complex contains a plant protein related to the RNA-binding subunit of yeast eIF-3. J Virol 71 6075 6082
36. FujisakiKRaveloGBNaitoSIshikawaM 2006 Involvement of THH1, an Arabidopsis thaliana homologue of the TOM1 gene, in tobamovirus multiplication. J Gen Virol 87 2397 2401
37. YamanakaTImaiTSatohRKawashimaATakahashiM 2002 Complete inhibition of tobamovirus multiplication by simultaneous mutations in two homologous host genes. J Virol 76 2491 2497
38. FujisakiKKobayashiSTsujimotoYNaitoSIshikawaM 2008 Analysis of tobamovirus multiplication in Arabidopsis thaliana mutants defective in TOM2A homologues. J Gen Virol 89 1519 1524
39. KomodaKNaitoSIshikawaM 2004 Replication of plant RNA virus genomes in a cell-free extract of evacuolated plant protoplasts. Proc Natl Acad Sci U S A 101 1863 1867
40. OkaiTArakiYTadaMTatenoTKontaniK 2004 Novel small GTPase subfamily capable of associating with tubulin is required for chromosome segregation. J Cell Sci 117 4705 4715
41. PasqualatoSRenaultLCherfilsJ 2002 Arf, Arl, Arp and Sar proteins: a family of GTP-binding proteins with a structural device for ‘front-back' communication. EMBO Rep 3 1035 1041
42. KahnRACherfilsJEliasMLoveringRCMunroS 2006 Nomenclature for the human Arf family of GTP-binding proteins: ARF, ARL, and SAR proteins. J Cell Biol 172 645 650
43. BagshawRDCallahanJWMahuranDJ 2006 The Arf-family protein, Arl8b, is involved in the spatial distribution of lysosomes. Biochem Biophys Res Commun 344 1186 1191
44. NakaeIFujinoTKobayashiTSasakiAKikkoY 2010 The arf-like GTPase Arl8 mediates delivery of endocytosed macromolecules to lysosomes in Caenorhabditis elegans. Mol Biol Cell 21 2434 2442
45. HofmannIMunroS 2006 An N-terminally acetylated Arf-like GTPase is localised to lysosomes and affects their motility. J Cell Sci 119 1494 1503
46. HeazlewoodJLVerboomRETonti-FilippiniJSmallIMillarAH 2007 SUBA: the Arabidopsis Subcellular Database. Nucleic acids res 35 D213 218
47. StagljarIKorostenskyCJohnssonNte HeesenS 1998 A genetic system based on split-ubiquitin for the analysis of interactions between membrane proteins in vivo. Proc Natl Acad Sci U S A 95 5187 5192
48. NielsenMAlbrethsenJLarsenFHSkriverK 2006 The Arabidopsis ADP-ribosylation factor (ARF) and ARF-like (ARL) system and its regulation by BIG2, a large ARF-GEF. Plant Science 171 707 717
49. NagyPD 2008 Yeast as a model host to explore plant virus-host interactions. Annu Rev Phytopathol 46 217 242
50. GraberJHMcAllisterGDSmithTF 2002 Probabilistic prediction of Saccharomyces cerevisiae mRNA 3′-processing sites. Nucleic Acids Res 30 1851 1858
51. MeritsAKettunenRMakinenKLampioAAuvinenP 1999 Virus-specific capping of tobacco mosaic virus RNA: methylation of GTP prior to formation of covalent complex p126-m7GMP. FEBS Lett 455 45 48
52. HuangYLHsuYHHanYTMengM 2005 mRNA guanylation catalyzed by the S-adenosylmethionine-dependent guanylyltransferase of bamboo mosaic virus. J Biol Chem 280 13153 13162
53. LiYIShihTWHsuYHHanYTHuangYL 2001 The helicase-like domain of plant potexvirus replicase participates in formation of RNA 5′ cap structure by exhibiting RNA 5′-triphosphatase activity. J Virol 75 12114 12120
54. AholaTden BoonJAAhlquistP 2000 Helicase and capping enzyme active site mutations in brome mosaic virus protein 1a cause defects in template recruitment, negative-strand RNA synthesis, and viral RNA capping. J Virol 74 8803 8811
55. WuSXAhlquistPKaesbergP 1992 Active complete in vitro replication of nodavirus RNA requires glycerophospholipid. Proc Natl Acad Sci U S A 89 11136 11140
56. den BoonJADiazAAhlquistP 2010 Cytoplasmic viral replication complexes. Cell Host Microbe 8 77 85
57. den BoonJAAhlquistP 2010 Organelle-like membrane compartmentalization of positive-strand RNA virus replication factories. Annu Rev Microbiol 64 241 256
58. LampioAKilpelainenIPesonenSKarhiKAuvinenP 2000 Membrane binding mechanism of an RNA virus-capping enzyme. J Biol Chem 275 37853 37859
59. LiuLWestlerWMden BoonJAWangXDiazA 2009 An amphipathic alpha-helix controls multiple roles of brome mosaic virus protein 1a in RNA replication complex assembly and function. PLoS Pathog 5 e1000351
60. MillerDJAhlquistP 2002 Flock house virus RNA polymerase is a transmembrane protein with amino-terminal sequences sufficient for mitochondrial localization and membrane insertion. J Virol 76 9856 9867
61. KubotaKTsudaSTamaiAMeshiT 2003 Tomato mosaic virus replication protein suppresses virus-targeted posttranscriptional gene silencing. J Virol 77 11016 11026
62. HagiwaraYKomodaKYamanakaTTamaiAMeshiT 2003 Subcellular localization of host and viral proteins associated with tobamovirus RNA replication. EMBO J 22 344 353
63. WangXGoregaokerSPCulverJN 2009 Interaction of the Tobacco mosaic virus replicase protein with a NAC domain transcription factor is associated with the suppression of systemic host defenses. J Virol 83 9720 9730
64. PadmanabhanMSGoregaokerSPGolemSShiferawHCulverJN 2005 Interaction of the tobacco mosaic virus replicase protein with the Aux/IAA protein PAP1/IAA26 is associated with disease development. J Virol 79 2549 2558
65. DohiKNishikioriMTamaiAIshikawaMMeshiT 2006 Inducible virus-mediated expression of a foreign protein in suspension-cultured plant cells. Arch Virol 151 1075 1084
66. MoriMKaidoMOkunoTFurusawaI 1993 mRNA amplification system by viral replicase in transgenic plants. FEBS Lett 336 171 174
67. OkamuraHNishikioriMXiangHIshikawaMKatohE 2011 Interconversion of two GDP-bound conformations and their selection in an Arf-family small G protein. Structure 19 988 998
68. ColicelliJBirchmeierCMichaeliTO'NeillKRiggsM 1989 Isolation and characterization of a mammalian gene encoding a high-affinity cAMP phosphodiesterase. Proc Natl Acad Sci U S A 86 3599 3603
69. GietzRDSuginoA 1988 New yeast-Escherichia coli shuttle vectors constructed with in vitro mutagenized yeast genes lacking six-base pair restriction sites. Gene 74 527 534
70. IshikawaMNaitoSOhnoT 1993 Effects of the tom1 mutation of Arabidopsis thaliana on the multiplication of tobacco mosaic virus RNA in protoplasts. J Virol 67 5328 5338
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2011 Číslo 12
- Parazitičtí červi v terapii Crohnovy choroby a dalších zánětlivých autoimunitních onemocnění
- Očkování proti virové hemoragické horečce Ebola experimentální vakcínou rVSVDG-ZEBOV-GP
- Koronavirus hýbe světem: Víte jak se chránit a jak postupovat v případě podezření?
Najčítanejšie v tomto čísle
- Controlling Viral Immuno-Inflammatory Lesions by Modulating Aryl Hydrocarbon Receptor Signaling
- Fungal Virulence and Development Is Regulated by Alternative Pre-mRNA 3′End Processing in
- Epstein-Barr Virus Nuclear Antigen 3C Stabilizes Gemin3 to Block p53-mediated Apoptosis
- Engineered Immunity to Infection