1887

Abstract

We have investigated the possible role of -acting factors interacting with the untranslated regions (UTRs) of coxsackievirus B3 (CVB3) RNA. We show here that polypyrimidine tract-binding protein (PTB) binds specifically to both 5′ and 3′ UTRs, but with different affinity. We have demonstrated that PTB is a bona fide internal ribosome entry site (IRES) -acting factor (ITAF) for CVB3 RNA by characterizing the effect of partial silencing of PTB in HeLa cells. Furthermore, IRES activity in BSC-1 cells, which are reported to have a very low level of endogenous PTB, was found to be significantly lower than that in HeLa cells. Additionally, we have mapped the putative contact points of PTB on the 5′ and 3′ UTRs by an RNA toe-printing assay. We have shown that the 3′ UTR is able to stimulate CVB3 IRES-mediated translation. Interestingly, a deletion of 15 nt at the 5′ end or 14 nt at the 3′ end of the CVB3 3′ UTR reduced the 3′ UTR-mediated enhancement of IRES activity significantly, and a reduced interaction was shown with PTB. It appears that the PTB protein might help in circularization of the CVB3 RNA by bridging the ends necessary for efficient translation of the viral RNA.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.018507-0
2010-05-01
2024-03-29
Loading full text...

Full text loading...

/deliver/fulltext/jgv/91/5/1245.html?itemId=/content/journal/jgv/10.1099/vir.0.018507-0&mimeType=html&fmt=ahah

References

  1. Baboonian, C., Davies, M. J., Booth, J. C. & Mckenna, W. J.(1997). Coxsackie B viruses and human heart disease. Curr Top Microbiol Immunol 223, 31–52. [Google Scholar]
  2. Bhattacharyya, S. & Das, S.(2005). Mapping of secondary structure of the spacer region within the 5′-untranslated region of the coxsackievirus B3 RNA: possible role of an apical GAGA loop in binding La protein and influencing internal initiation of translation. Virus Res 108, 89–100.[CrossRef] [Google Scholar]
  3. Bhattacharyya, S. & Das, S.(2006). An apical GAGA loop within 5′-UTR of the coxsackievirus B3 RNA maintains structural organization of the IRES element required for efficient ribosome entry. RNA Biol 3, 60–68.[CrossRef] [Google Scholar]
  4. Bhattacharyya, S., Verma, B., Pandey, G. & Das, S.(2008). The structure and function of a cis-acting element located upstream of the IRES that influences coxsackievirus B3 RNA translation. Virology 377, 345–354.[CrossRef] [Google Scholar]
  5. Chen, T. C., Weng, K. F., Chang, S. C., Lin, J. Y., Huang, P. N. & Shih, S. R.(2008). Development of antiviral agents for enteroviruses. J Antimicrob Chemother 62, 1169–1173.[CrossRef] [Google Scholar]
  6. Dhar, D., Roy, S. & And Das, S.(2007). Translational control of the interferon regulatory factor 2 mRNA by IRES element. Nucleic Acids Res 35, 5409–5421.[CrossRef] [Google Scholar]
  7. Dobrikova, E. Y., Grisham, R. N., Kaiser, C., Lin, J. & Gromeier, M.(2006). Competitive translation efficiency at the picornavirus type 1 internal ribosome entry site facilitated by viral cis and trans factors. J Virol 80, 3310–3321.[CrossRef] [Google Scholar]
  8. Fitzgerald, K. D. & Semler, B. L.(2009). Bridging IRES elements in mRNAs to the eukaryotic translation apparatus. Biochim Biophys Acta 1789, 518–528.[CrossRef] [Google Scholar]
  9. Gosert, R., Chang, K. H., Rijnbrand, R., Yi, M., Sangar, D. V. & Lemon, S. M.(2000). Transient expression of cellular polypyrimidine-tract binding protein stimulates cap-independent translation directed by both picornaviral and flaviviral internal ribosome entry sites in vivo. Mol Cell Biol 20, 1583–1595.[CrossRef] [Google Scholar]
  10. Grover, R., Ray, P. S. & Das, S.(2008). Polypyrimidine tract binding protein regulates IRES-mediated translation of p53 isoforms. Cell Cycle 7, 2189–2198.[CrossRef] [Google Scholar]
  11. Hellen, C. U., Pestova, T. V., Litterst, M. & Wimmer, E.(1994). The cellular polypeptide p57 (pyrimidine tract-binding protein) binds to multiple sites in the poliovirus 5′ nontranslated region. J Virol 68, 941–950. [Google Scholar]
  12. Hunt, S. L. & Jackson, R.(1999). Polypyrimidine-tract binding protein (PTB) is necessary, but not sufficient, for efficient internal initiation of translation of human rhinovirus-2 RNA. RNA 5, 344–359.[CrossRef] [Google Scholar]
  13. Jang, S. K., Kräusslich, H. G., Nicklin, M. J., Duke, G. M., Palmenberg, A. C. & Wimmer, E.(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. [Google Scholar]
  14. Kafasla, P., Morgner, N., Pöyry, T. A., Curry, S., Robinson, C. V. & Jackson, R. J.(2009). Polypyrimidine tract binding protein stabilizes the encephalomyocarditis virus IRES structure via binding multiple sites in a unique orientation. Mol Cell 34, 556–568.[CrossRef] [Google Scholar]
  15. Klump, W. M., Bergmann, I., Müller, B. C., Ameis, D. & Kandolf, R.(1990). Complete nucleotide sequence of infectious coxsackievirus B3 cDNA: two initial 5′ uridine residues are regained during plus-strand RNA synthesis. J Virol 64, 1573–1583. [Google Scholar]
  16. Kolupaeva, V. G., Hellen, C. U. & Shatsky, I. N.(1996). Structural analysis of the interaction of the pyrimidine tract-binding protein with the internal ribosomal entry site of encephalomyocarditis virus and foot-and-mouth disease virus RNAs. RNA 2, 1199–1212. [Google Scholar]
  17. Melchers, W. J. G., Hoenderop, J. G. J., Bruins Slot, H. J., Pleij, C. W. A., Pilipenko, E. V., Agol, V. I. & Galama, J. M. D.(1997). Kissing of the two predominant hairpin loops in the coxsackie B virus 3′ untranslated region is the essential structural feature of the origin of replication required for negative-strand RNA synthesis. J Virol 71, 686–696. [Google Scholar]
  18. Merkle, I., van Ooij, M. J. M., van Kuppeveld, F. J. M., Glaudemans, D. H. R. F., Galama, J. M. D., Henke, A., Zell, R. & Melchers, W. J. G.(2002). Biological significance of a human enterovirus B-specific RNA element in the 3′ nontranslated region. J Virol 76, 9900–9909.[CrossRef] [Google Scholar]
  19. M'hadheb-Gharbi, M. B., Paulous, S., Aouni, M., Kean, K. M. & Gharbi, J.(2007). The substitution U475→C with Sabin3-like mutation within the IRES attenuate coxsackievirus B3 cardiovirulence. Mol Biotechnol 36, 52–60.[CrossRef] [Google Scholar]
  20. Niepmann, M., Petersen, A., Meyer, K. & Beck, E.(1997). Functional involvement of polypyrimidine tract-binding protein in translation initiation complexes with the internal ribosome entry site of foot-and-mouth disease virus. J Virol 71, 8330–8339. [Google Scholar]
  21. Pestova, T. V., Hellen, C. U. T. & Wimmer, E.(1991). Translation of poliovirus RNA: role of an essential cis-acting oligopyrimidine element within the 5′ nontranslated region and involvement of a cellular 57-kilodalton protein. J Virol 65, 6194–6204. [Google Scholar]
  22. Ray, P. S. & Das, S.(2002). La autoantigen is required for the internal ribosome entry site-mediated translation of coxsackievirus B3 RNA. Nucleic Acids Res 30, 4500–4508.[CrossRef] [Google Scholar]
  23. Schultz, D. E., Hardin, C. C. & Lemon, S. M.(1996). Specific interaction of glyceraldehyde 3-phosphate dehydrogenase with the 5′-nontranslated RNA of hepatitis A virus. J Biol Chem 271, 14134–14142.[CrossRef] [Google Scholar]
  24. Serrano, P., Pulido, M. R., Saiz, M. & Martinez-Salas, E.(2006). The 3′ end of the foot-and-mouth disease virus genome establishes two distinct long-range RNA–RNA interactions with the 5′ end region. J Gen Virol 87, 3013–3022.[CrossRef] [Google Scholar]
  25. Song, Y., Tzima, E., Ochs, K., Bassili, G., Trusheim, H., Linder, M., Preissner, K. T. & Niepmann, M.(2005). Evidence for an RNA chaperone function of polypyrimidine tract-binding protein in picornavirus translation. RNA 11, 1809–1824.[CrossRef] [Google Scholar]
  26. Song, Y., Friebe, P., Tzima, E., Junemann, C., Bartenschlager, R. & Niepmann, M.(2006). The hepatitis C virus RNA 3′-untranslated region strongly enhances translation directed by the internal ribosome entry site. J Virol 80, 11579–11588.[CrossRef] [Google Scholar]
  27. Vagner, S., Galy, B. & Pyronnet, S.(2001). Irresistible IRES. Attracting the translation machinery to internal ribosome entry sites. EMBO Rep 2, 893–898.[CrossRef] [Google Scholar]
  28. Venkatramana, M., Ray, P. S., Chadda, A. & Das, S.(2003). A 25 kDa cleavage product of polypyrimidine tract binding protein (PTB) present in mouse tissues prevents PTB binding to the 5′ untranslated region and inhibits translation of hepatitis A virus RNA. Virus Res 98, 141–149.[CrossRef] [Google Scholar]
  29. Wagner, E. J. & Garcia-Blanco, M. A.(2002). RNAi-mediated depletion of PTB leads to enhanced exon definition. Mol Cell 10, 943–949.[CrossRef] [Google Scholar]
  30. Zell, R., Ihle, Y., Seitz, S., Gündel, U., Wutzler, P. & Görlach, M.(2008). Poly(rC)-binding protein 2 interacts with the oligo(rC) tract of coxsackievirus B3. Biochem Biophys Res Commun 366, 917–921.[CrossRef] [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.018507-0
Loading
/content/journal/jgv/10.1099/vir.0.018507-0
Loading

Data & Media loading...

This is a required field
Please enter a valid email address
Approval was a Success
Invalid data
An Error Occurred
Approval was partially successful, following selected items could not be processed due to error