1887

Abstract

Enterovirus 71 (EV71) is a picornavirus that can cause severe neurological complications in children. Like other picornaviruses, the genomic RNA of EV71 contains a long 5′ untranslated region (UTR). Cellular proteins interact with the EV71 5′ UTR, and these interactions are important for virus replication. Using an RNA pull-down assay and proteomics approaches, this study identified the heterogeneous nuclear ribonucleoprotein K (hnRNP K) as one of the EV71 5′ UTR-associated proteins. The interaction between hnRNP K and the 5′ UTR was further confirmed by mapping the interaction regions to stem–loops I–II and IV in the 5′ UTR. During EV71 infection, hnRNP K was enriched in the cytoplasm where virus replication occurs, whereas hnRNP K was localized in the nucleus in mock-infected cells. Viral yields were found to be significantly lower in hnRNP K knockdown cells and viral RNA synthesis was delayed in hnRNP K knockdown cells in comparison with negative-control cells treated with small interfering RNA. These results suggest that hnRNP K interacts with the EV71 5′ UTR and participates in virus replication.

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2008-10-01
2020-04-03
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References

  1. Adinolfi S., Bagni C., Castiglione Morelli M. A., Fraternali F., Musco G., Pastore A. 1999; Novel RNA-binding motif: the KH module. Biopolymers 51:153–164 [CrossRef]
    [Google Scholar]
  2. Bomsztyk K., Denisenko O., Ostrowski J. 2004; hnRNP K: one protein multiple processes. Bioessays 26:629–638 [CrossRef]
    [Google Scholar]
  3. Boussadia O., Niepmann M., Creancier L., Prats A. C., Dautry F., Jacquemin-Sablon H. 2003; Unr is required in vivo for efficient initiation of translation from the internal ribosome entry sites of both rhinovirus and poliovirus. J Virol 77:3353–3359 [CrossRef]
    [Google Scholar]
  4. Brown B. A., Pallansch M. A. 1995; Complete nucleotide sequence of enterovirus 71 is distinct from poliovirus. Virus Res 39:195–205 [CrossRef]
    [Google Scholar]
  5. Brunner J. E., Nguyen J. H., Roehl H. H., Ho T. V., Swiderek K. M., Semler B. L. 2005; Functional interaction of heterogeneous nuclear ribonucleoprotein C with poliovirus RNA synthesis initiation complexes. J Virol 79:3254–3266 [CrossRef]
    [Google Scholar]
  6. Bryant H. E., Matthews D. A., Wadd S., Scott J. E., Kean J., Graham S., Russell W. C., Clements J. B. 2000; Interaction between herpes simplex virus type 1 IE63 protein and cellular protein p32. J Virol 74:11322–11328 [CrossRef]
    [Google Scholar]
  7. Burnham A. J., Gong L., Hardy R. W. 2007; Heterogeneous nuclear ribonuclear protein K interacts with Sindbis virus nonstructural proteins and viral subgenomic mRNA. Virology 367:212–221 [CrossRef]
    [Google Scholar]
  8. Chang C. J., Luh H. W., Wang S. H., Lin H. J., Lee S. C., Hu S. T. 2001; The heterogeneous nuclear ribonucleoprotein K (hnRNP K) interacts with dengue virus core protein. DNA Cell Biol 20:569–577 [CrossRef]
    [Google Scholar]
  9. Chen K. T., Chang H. L., Wang S. T., Cheng Y. T., Yang J. Y. 2007; Epidemiologic features of hand-foot-mouth disease and herpangina caused by enterovirus 71 in Taiwan, 1998–2005. Pediatrics 120:e244–e252 [CrossRef]
    [Google Scholar]
  10. Choi K., Kim J. H., Li X., Paek K. Y., Ha S. H., Ryu S. H., Wimmer E., Jang S. K. 2004; Identification of cellular proteins enhancing activities of internal ribosomal entry sites by competition with oligodeoxynucleotides. Nucleic Acids Res 32:1308–1317 [CrossRef]
    [Google Scholar]
  11. Collier B., Goobar-Larsson L., Sokolowski M., Schwartz S. 1998; Translational inhibition in vitro of human papillomavirus type 16 L2 mRNA mediated through interaction with heterogenous ribonucleoprotein K and poly(rC)-binding proteins 1 and 2. J Biol Chem 273:22648–22656 [CrossRef]
    [Google Scholar]
  12. Costa-Mattioli M., Svitkin Y., Sonenberg N. 2004; La autoantigen is necessary for optimal function of the poliovirus and hepatitis C virus internal ribosome entry site in vivo and in vitro. Mol Cell Biol 24:6861–6870 [CrossRef]
    [Google Scholar]
  13. Dejgaard K., Leffers H. 1996; Characterisation of the nucleic-acid-binding activity of KH domains. Different properties of different domains. Eur J Biochem 241:425–431 [CrossRef]
    [Google Scholar]
  14. Evans J. R., Mitchell S. A., Spriggs K. A., Ostrowski J., Bomsztyk K., Ostarek D., Willis A. E. 2003; Members of the poly (rC) binding protein family stimulate the activity of the c- myc internal ribosome entry segment in vitro and in vivo . Oncogene 22:8012–8020 [CrossRef]
    [Google Scholar]
  15. Flanegan J. B., Petterson R. F., Ambros V., Hewlett N. J., Baltimore D. 1977; Covalent linkage of a protein to a defined nucleotide sequence at the 5′-terminus of virion and replicative intermediate RNAs of poliovirus. Proc Natl Acad Sci U S A 74:961–965 [CrossRef]
    [Google Scholar]
  16. Gibson T. J., Thompson J. D., Heringa J. 1993; The KH domain occurs in a diverse set of RNA-binding proteins that include the antiterminator NusA and is probably involved in binding to nucleic acid. FEBS Lett 324:361–366 [CrossRef]
    [Google Scholar]
  17. Herold J., Andino R. 2001; Poliovirus RNA replication requires genome circularization through a protein–protein bridge. Mol Cell 7:581–591 [CrossRef]
    [Google Scholar]
  18. Ho M., Chen E. R., Hsu K. H., Twu S. J., Chen K. T., Tsai S. F., Wang J. R., Shih S. R. 1999; An epidemic of enterovirus 71 infection in Taiwan. Taiwan Enterovirus Epidemic Working Group. N Engl J Med 341:929–935 [CrossRef]
    [Google Scholar]
  19. Hsieh T. Y., Matsumoto M., Chou H. C., Schneider R., Hwang S. B., Lee A. S., Lai M. M. 1998; Hepatitis C virus core protein interacts with heterogeneous nuclear ribonucleoprotein K. J Biol Chem 273:17651–17659 [CrossRef]
    [Google Scholar]
  20. Inoue A., Sawata S. Y., Taira K., Wadhwa R. 2007; Loss-of-function screening by randomized intracellular antibodies: identification of hnRNP-K as a potential target for metastasis. Proc Natl Acad Sci U S A 104:8983–8988 [CrossRef]
    [Google Scholar]
  21. Lee P. T., Liao P. C., Chang W. C., Tseng J. T. 2007; Epidermal growth factor increases the interaction between nucleolin and heterogeneous nuclear ribonucleoprotein K/poly(C) binding protein 1 complex to regulate the gastrin mRNA turnover. Mol Biol Cell 18:5004–5013 [CrossRef]
    [Google Scholar]
  22. Leopoldino A. M., Carregaro F., Silva C. H., Feitosa O., Mancini U. M., Freitas J. M., Tajara E. H. 2007; Sequence and transcriptional study of HNRPK pseudogenes, and expression and molecular modeling analysis of hnRNP K isoforms. Genome 50:451–462 [CrossRef]
    [Google Scholar]
  23. McMinn P. C. 2002; An overview of the evolution of enterovirus 71 and its clinical and public health significance. FEMS Microbiol Rev 26:91–107 [CrossRef]
    [Google Scholar]
  24. McMinn P., Lindsay K., Perera D., Chan H. M., Chan K. P., Cardosa M. J. 2001; Phylogenetic analysis of enterovirus 71 strains isolated during linked epidemics in Malaysia, Singapore, and Western Australia. J Virol 75:7732–7738 [CrossRef]
    [Google Scholar]
  25. Messias A. C., Sattler M. 2004; Structural basis of single-stranded RNA recognition. Acc Chem Res 37:279–287 [CrossRef]
    [Google Scholar]
  26. Murray K. E., Roberts A. W., Barton D. J. 2001; Poly(rC) binding proteins mediate poliovirus mRNA stability. RNA 7:1126–1141 [CrossRef]
    [Google Scholar]
  27. Parsley T. B., Towner J. S., Blyn L. B., Ehrenfeld E., Semler B. L. 1997; Poly(rC) binding protein 2 forms a ternary complex with the 5′-terminal sequences of poliovirus RNA and the viral 3CD proteinase. RNA 3:1124–1134
    [Google Scholar]
  28. Sarnow P. 1989; Role of 3′-end sequences in infectivity of poliovirus transcripts made in vitro. J Virol 63:467–470
    [Google Scholar]
  29. Shimada K., Kondo K., Yamanishi K. 2004; Human herpesvirus 6 immediate-early 2 protein interacts with heterogeneous ribonucleoprotein K and casein kinase 2. Microbiol Immunol 48:205–210 [CrossRef]
    [Google Scholar]
  30. Silvera D., Gamarnik A. V., Andino R. 1999; The N-terminal K homology domain of the poly(rC)-binding protein is a major determinant for binding to the poliovirus 5′-untranslated region and acts as an inhibitor of viral translation. J Biol Chem 274:38163–38170 [CrossRef]
    [Google Scholar]
  31. Siomi H., Matunis M. J., Michael W. M., Dreyfuss G. 1993; The pre-mRNA binding K protein contains a novel evolutionarily conserved motif. Nucleic Acids Res 21:1193–1198 [CrossRef]
    [Google Scholar]
  32. Thompson S. R., Sarnow P. 2003; Enterovirus 71 contains a type I IRES element that functions when eukaryotic initiation factor eIF4G is cleaved. Virology 315:259–266 [CrossRef]
    [Google Scholar]
  33. Tomonaga T., Levens D. 1995; Heterogeneous nuclear ribonucleoprotein K is a DNA-binding transactivator. J Biol Chem 270:4875–4881 [CrossRef]
    [Google Scholar]
  34. Toyoda H., Koide N., Kamiyama M., Tobita K., Mizumoto K., Imura N. 1994; Host factors required for internal initiation of translation on poliovirus RNA. Arch Virol 138:1–15 [CrossRef]
    [Google Scholar]
  35. Walter B. L., Nguyen J. H., Ehrenfeld E., Semler B. L. 1999; Differential utilization of poly(rC) binding protein 2 in translation directed by picornavirus IRES elements. RNA 5:1570–1585 [CrossRef]
    [Google Scholar]
  36. Walter B. L., Parsley T. B., Ehrenfeld E., Semler B. L. 2002; Distinct poly(rC) binding protein KH domain determinants for poliovirus translation initiation and viral RNA replication. J Virol 76:12008–12022 [CrossRef]
    [Google Scholar]
  37. Wang Z., Day N., Trifillis P., Kiledjian M. 1999; An mRNA stability complex functions with poly(A)-binding protein to stabilize mRNA in vitro. Mol Cell Biol 19:4552–4560
    [Google Scholar]
  38. Zhang W., Zhang X., Tian C., Wang T., Sarkis P. T., Fang Y., Zheng S., Yu X. F., Xu R. 2008; Cytidine deaminase APOBEC3B interacts with heterogeneous nuclear ribonucleoprotein K and suppresses hepatitis B virus expression. Cell Microbiol 10:112–121
    [Google Scholar]
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