Re-localization of Cellular Protein SRp20 during Poliovirus Infection: Bridging a Viral IRES to the Host Cell Translation Apparatus
Poliovirus IRES-mediated translation requires the functions of certain canonical as well as non-canonical factors for the recruitment of ribosomes to the viral RNA. The interaction of cellular proteins PCBP2 and SRp20 in extracts from poliovirus-infected cells has been previously described, and these two proteins were shown to function synergistically in viral translation. To further define the mechanism of ribosome recruitment for the initiation of poliovirus IRES-dependent translation, we focused on the role of the interaction between cellular proteins PCBP2 and SRp20. Work described here demonstrates that SRp20 dramatically re-localizes from the nucleus to the cytoplasm of poliovirus-infected neuroblastoma cells during the course of infection. Importantly, SRp20 partially co-localizes with PCBP2 in the cytoplasm of infected cells, corroborating our previous in vitro interaction data. In addition, the data presented implicate the presence of these two proteins in viral translation initiation complexes. We show that in extracts from poliovirus-infected cells, SRp20 is associated with PCBP2 bound to poliovirus RNA, indicating that this interaction occurs on the viral RNA. Finally, we generated a mutated version of SRp20 lacking the RNA recognition motif (SRp20ΔRRM) and found that this protein is localized similar to the full length SRp20, and also partially co-localizes with PCBP2 during poliovirus infection. Expression of this mutated version of SRp20 results in a ∼100 fold decrease in virus yield for poliovirus when compared to expression of wild type SRp20, possibly via a dominant negative effect. Taken together, these results are consistent with a model in which SRp20 interacts with PCBP2 bound to the viral RNA, and this interaction functions to recruit ribosomes to the viral RNA in a direct or indirect manner, with the participation of additional protein-protein or protein-RNA interactions.
Vyšlo v časopise:
Re-localization of Cellular Protein SRp20 during Poliovirus Infection: Bridging a Viral IRES to the Host Cell Translation Apparatus. PLoS Pathog 7(7): e32767. doi:10.1371/journal.ppat.1002127
Kategorie:
Research Article
prolekare.web.journal.doi_sk:
https://doi.org/10.1371/journal.ppat.1002127
Souhrn
Poliovirus IRES-mediated translation requires the functions of certain canonical as well as non-canonical factors for the recruitment of ribosomes to the viral RNA. The interaction of cellular proteins PCBP2 and SRp20 in extracts from poliovirus-infected cells has been previously described, and these two proteins were shown to function synergistically in viral translation. To further define the mechanism of ribosome recruitment for the initiation of poliovirus IRES-dependent translation, we focused on the role of the interaction between cellular proteins PCBP2 and SRp20. Work described here demonstrates that SRp20 dramatically re-localizes from the nucleus to the cytoplasm of poliovirus-infected neuroblastoma cells during the course of infection. Importantly, SRp20 partially co-localizes with PCBP2 in the cytoplasm of infected cells, corroborating our previous in vitro interaction data. In addition, the data presented implicate the presence of these two proteins in viral translation initiation complexes. We show that in extracts from poliovirus-infected cells, SRp20 is associated with PCBP2 bound to poliovirus RNA, indicating that this interaction occurs on the viral RNA. Finally, we generated a mutated version of SRp20 lacking the RNA recognition motif (SRp20ΔRRM) and found that this protein is localized similar to the full length SRp20, and also partially co-localizes with PCBP2 during poliovirus infection. Expression of this mutated version of SRp20 results in a ∼100 fold decrease in virus yield for poliovirus when compared to expression of wild type SRp20, possibly via a dominant negative effect. Taken together, these results are consistent with a model in which SRp20 interacts with PCBP2 bound to the viral RNA, and this interaction functions to recruit ribosomes to the viral RNA in a direct or indirect manner, with the participation of additional protein-protein or protein-RNA interactions.
Zdroje
1. KozakM 1984 Compilation and analysis of sequences upstream from the translational start site in eukaryotic mRNAs. Nucleic Acids Res 12 857 872
2. FitzgeraldKDSemlerBL 2009 Bridging IRES elements in mRNAs to the eukaryotic translation apparatus. Biochim Biophys Acta 1789 518 528
3. BairdSDLewisSMTurcotteMHolcikM 2007 A search for structurally similar cellular internal ribosome entry sites. Nucleic Acids Res 35 4664 4677
4. BairdSDTurcotteMKornelukRGHolcikM 2006 Searching for IRES. RNA 12 1755 1785
5. JangSKKrausslichHGNicklinMJDukeGMPalmenbergAC 1988 A segment of the 5′ nontranslated region of encephalomyocarditis virus RNA directs internal entry of ribosomes during in vitro translation. J Virol 62 2636 2643
6. PelletierJKaplanGRacanielloVRSonenbergN 1988 Cap-independent translation of poliovirus mRNA is conferred by sequence elements within the 5′ noncoding region. Mol Cell Biol 8 1103 1112
7. PelletierJSonenbergN 1988 Internal initiation of translation of eukaryotic mRNA directed by a sequence derived from poliovirus RNA. Nature 334 320 325
8. EtchisonDMilburnSCEderyISonenbergNHersheyJW 1982 Inhibition of HeLa cell protein synthesis following poliovirus infection correlates with the proteolysis of a 220,000-dalton polypeptide associated with eucaryotic initiation factor 3 and a cap binding protein complex. J Biol Chem 257 14806 14810
9. KrausslichHGNicklinMJToyodaHEtchisonDWimmerE 1987 Poliovirus proteinase 2A induces cleavage of eucaryotic initiation factor 4F polypeptide p220. J Virol 61 2711 2718
10. LloydREJenseHGEhrenfeldE 1987 Restriction of translation of capped mRNA in vitro as a model for poliovirus-induced inhibition of host cell protein synthesis: relationship to p220 cleavage. J Virol 61 2480 2488
11. JoachimsMVan BreugelPCLloydRE 1999 Cleavage of poly(A)-binding protein by enterovirus proteases concurrent with inhibition of translation in vitro. J Virol 73 718 727
12. KerekatteVKeiperBDBadorffCCaiAKnowltonKU 1999 Cleavage of Poly(A)-binding protein by coxsackievirus 2A protease in vitro and in vivo: another mechanism for host protein synthesis shutoff? J Virol 73 709 717
13. Kuyumcu-MartinezNMJoachimsMLloydRE 2002 Efficient cleavage of ribosome-associated poly(A)-binding protein by enterovirus 3C protease. J Virol 76 2062 2074
14. MakeyevAVLiebhaberSA 2002 The poly(C)-binding proteins: a multiplicity of functions and a search for mechanisms. RNA 8 265 278
15. WalterBLNguyenJHEhrenfeldESemlerBL 1999 Differential utilization of poly(rC) binding protein 2 in translation directed by picornavirus IRES elements. RNA 5 1570 1585
16. BlynLBTownerJSSemlerBLEhrenfeldE 1997 Requirement of poly(rC) binding protein 2 for translation of poliovirus RNA. J Virol 71 6243 6246
17. WalterBLParsleyTBEhrenfeldESemlerBL 2002 Distinct poly(rC) binding protein KH domain determinants for poliovirus translation initiation and viral RNA replication. J Virol 76 12008 12022
18. BedardKMDaijogoSSemlerBL 2007 A nucleo-cytoplasmic SR protein functions in viral IRES-mediated translation initiation. EMBO J 26 459 467
19. ZahlerAMLaneWSStolkJARothMB 1992 SR proteins: a conserved family of pre-mRNA splicing factors. Genes Dev 6 837 847
20. CaceresJFMisteliTScreatonGRSpectorDLKrainerAR 1997 Role of the modular domains of SR proteins in subnuclear localization and alternative splicing specificity. J Cell Biol 138 225 238
21. CaceresJFScreatonGRKrainerAR 1998 A specific subset of SR proteins shuttles continuously between the nucleus and the cytoplasm. Genes Dev 12 55 66
22. HuangYSteitzJA 2001 Splicing factors SRp20 and 9G8 promote the nucleocytoplasmic export of mRNA. Mol Cell 7 899 905
23. LongJCCaceresJF 2009 The SR protein family of splicing factors: master regulators of gene expression. Biochem J 417 15 27
24. BjorkPJinSZhaoJSinghOPPerssonJO 2009 Specific combinations of SR proteins associate with single pre-messenger RNAs in vivo and contribute different functions. J Cell Biol 184 555 568
25. SanfordJRGrayNKBeckmannKCaceresJF 2004 A novel role for shuttling SR proteins in mRNA translation. Genes Dev 18 755 768
26. SwartzJEBorYCMisawaYRekoshDHammarskjoldML 2007 The shuttling SR protein 9G8 plays a role in translation of unspliced mRNA containing a constitutive transport element. J Biol Chem 282 19844 19853
27. SwansonCMShererNMMalimMH 2010 SRp40 and SRp55 promote the translation of unspliced human immunodeficiency virus type 1 RNA. J Virol 84 6748 6759
28. BaltimoreDGirardMDarnellJE 1966 Aspects of Synthesis of Poliovirus Rna and Formation of Virus Particles. Virology 29 179 189
29. PuckTTMarcusPICieciuraSJ 1956 Clonal growth of mammalian cells in vitro; growth characteristics of colonies from single HeLa cells with and without a feeder layer. J Exp Med 103 273 283
30. HargousYHautbergueGMTintaruAMSkrisovskaLGolovanovAP 2006 Molecular basis of RNA recognition and TAP binding by the SR proteins SRp20 and 9G8. EMBO J 25 5126 5137
31. KuhnRJWimmerESemlerBL 1987 Expression of the poliovirus genome from infectious cDNA is dependent upon arrangements of eukaryotic and prokaryotic sequences in recombinant plasmids. Virology 157 560 564
32. DildineSLSemlerBL 1992 Conservation of RNA-protein interactions among picornaviruses. J Virol 66 4364 4376
33. SeanPNguyenJHSemlerBL 2008 The linker domain of poly(rC) binding protein 2 is a major determinant in poliovirus cap-independent translation. Virology 378 243 253
34. NeugebauerKMRothMB 1997 Distribution of pre-mRNA splicing factors at sites of RNA polymerase II transcription. Genes Dev 11 1148 1159
35. GustinKESarnowP 2001 Effects of poliovirus infection on nucleo-cytoplasmic trafficking and nuclear pore complex composition. EMBO J 20 240 249
36. Escudero-PaunettoLLiLHernandezFPSandri-GoldinRM 2010 SR proteins SRp20 and 9G8 contribute to efficient export of herpes simplex virus 1 mRNAs. Virology 401 155 164
37. GamarnikAVAndinoR 1997 Two functional complexes formed by KH domain containing proteins with the 5′ noncoding region of poliovirus RNA. RNA 3 882 892
38. FujimuraKKanoFMurataM 2008 Identification of PCBP2, a facilitator of IRES-mediated translation, as a novel constituent of stress granules and processing bodies. RNA 14 425 431
39. SvitkinYVCosta-MattioliMHerdyBPerreaultSSonenbergN 2007 Stimulation of picornavirus replication by the poly(A) tail in a cell-free extract is largely independent of the poly(A) binding protein (PABP). RNA 13 2330 2340
40. SvitkinYVImatakaHKhaleghpourKKahvejianALiebigHD 2001 Poly(A)-binding protein interaction with elF4G stimulates picornavirus IRES-dependent translation. RNA 7 1743 1752
41. SummersDFMaizelJVJDarnellJEJ 1967 The decrease in size and synthetic activity of poliovirus polysomes late in the infectious cycle. Virology 31 427 435
42. JohannesGSarnowP 1998 Cap-independent polysomal association of natural mRNAs encoding c-myc, BiP, and eIF4G conferred by internal ribosome entry sites. RNA 4 1500 1513
43. JohannesGCarterMSEisenMBBrownPOSarnowP 1999 Identification of eukaryotic mRNAs that are translated at reduced cap binding complex eIF4F concentrations using a cDNA microarray. Proc Natl Acad Sci U S A 96 13118 13123
44. BlynLBSwiderekKMRichardsOStahlDCSemlerBL 1996 Poly(rC) binding protein 2 binds to stem-loop IV of the poliovirus RNA 5′ noncoding region: identification by automated liquid chromatography-tandem mass spectrometry. Proc Natl Acad Sci USA 93 11115 11120
45. BedardKMWalterBLSemlerBL 2004 Multimerization of poly(rC) binding protein 2 is required for translation initiation mediated by a viral IRES. RNA 10 1266 1276
46. KataokaNBachorikJLDreyfussG 1999 Transportin-SR, a nuclear import receptor for SR proteins. J Cell Biol 145 1145 1152
47. LaiMCLinRIHuangSYTsaiCWTarnWY 2000 A human importin-beta family protein, transportin-SR2, interacts with the phosphorylated RS domain of SR proteins. J Biol Chem 275 7950 7957
48. AllemandEDokudovskayaSBordonneRTaziJ 2002 A conserved Drosophila transportin-serine/arginine-rich (SR) protein permits nuclear import of Drosophila SR protein splicing factors and their antagonist repressor splicing factor 1. Mol Biol Cell 13 2436 2447
49. MeerovitchKSvitkinYVLeeHSLejbkowiczFKenanDJ 1993 La autoantigen enhances and corrects aberrant translation of poliovirus RNA in reticulocyte lysate. J Virol 67 3798 3807
50. BrunnerJEErtelKJRozovicsJMSemlerBL 2010 Delayed kinetics of poliovirus RNA synthesis in a human cell line with reduced levels of hnRNP C proteins. Virology 400 240 247
51. McBrideAESchlegelAKirkegaardK 1996 Human protein Sam68 relocalization and interaction with poliovirus RNA polymerase in infected cells. Proc Natl Acad Sci USA 93 2296 2301
52. ShirokiKIsoyamaTKugeSIshiiTOhmiS 1999 Intracellular redistribution of truncated La protein produced by poliovirus 3Cpro-mediated cleavage. J Virol 73 2193 2200
53. WaggonerSSarnowP 1998 Viral ribonucleoprotein complex formation and nucleolar-cytoplasmic relocalization of nucleolin in poliovirus-infected cells. J Virol 72 6699 6709
54. BackSHKimYKKimWJChoSOhHR 2002 Translation of polioviral mRNA is inhibited by cleavage of polypyrimidine tract-binding proteins executed by polioviral 3C(pro). J Virol 76 2529 2542
55. ParkNSkernTGustinKE 2010 Specific cleavage of the nuclear pore complex protein Nup62 by a viral protease. J Biol Chem 285 28796 28805
56. BelovGALidskyPVMikitasOVEggerDLukyanovKA 2004 Bidirectional increase in permeability of nuclear envelope upon poliovirus infection and accompanying alterations of nuclear pores. J Virol 78 10166 10177
57. SonenbergNHinnebuschAG 2009 Regulation of translation initiation in eukaryotes: mechanisms and biological targets. Cell 136 731 745
58. BlynLBChenRSemlerBLEhrenfeldE 1995 Host cell proteins binding to domain IV of the 5′ noncoding region of poliovirus RNA. J Virol 69 4381 4389
Štítky
Hygiena a epidemiológia Infekčné lekárstvo LaboratóriumČlánok vyšiel v časopise
PLOS Pathogens
2011 Číslo 7
- 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
- Requires Glycerol for Maximum Fitness During The Tick Phase of the Enzootic Cycle
- Comparative Genomics Yields Insights into Niche Adaptation of Plant Vascular Wilt Pathogens
- The Role of IL-15 Deficiency in the Pathogenesis of Virus-Induced Asthma Exacerbations
- “Persisters”: Survival at the Cellular Level