1887

Abstract

Polypeptide 2C is essential for picornavirus replication. Although many data on multiple functions of this highly conserved protein are available, the mechanism of RNA binding is still obscure. In this work, protein 2C of echovirus-9 strain Barty was expressed as a histidine-tagged protein in followed by nondenaturing purification to homogeneity. After incubation of 2C protein with different kinds of RNA fragments, binding was shown in gel retardation assays. Competition experiments revealed that 2C targets linear RNA unspecifically; however, single-stranded linear DNA does not react with this protein. In contrast to poliovirus, protein 2C of echovirus-9 only recognizes RNA with a low content of secondary structures. This may be a first hint of a different binding specificity of 2C in echo- and polioviruses.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-81-10-2481
2000-10-01
2024-04-26
Loading full text...

Full text loading...

/deliver/fulltext/jgv/81/10/0812481a.html?itemId=/content/journal/jgv/10.1099/0022-1317-81-10-2481&mimeType=html&fmt=ahah

References

  1. Argos P., Kamer G., Nicklin M. J. H., Wimmer E. 1984; Similarity in the gene organization and homology between proteins of animal picornaviruses and a plant comovirus suggest common ancestry of these virus families. Nucleic Acids Research 12:7251–7267
    [Google Scholar]
  2. Ausubel F. M., Brent R., Kingston R. E., Moore D. D., Seidmann J. G., Smith J. A., Struhl K. 1998 Current Protocols in Molecular Biology New York: John Wiley;
  3. Banerjee R., Echeverri A., Dasgupta A. 1997; Poliovirus-encoded 2C polypeptide specifically binds to the 3′-terminal sequences of viral negative-strand RNA. Journal of Virology 71:9570–9578
    [Google Scholar]
  4. Bienz K., Egger D., Pfister T., Troxler M. 1992; Structural and functional characterization of the poliovirus replication complex. Journal of Virology 66:2740–2747
    [Google Scholar]
  5. Citovsky V., Wong M. L., Shaw A. L., Prasad B. V. V., Zambryski P. 1992; Visualization and characterisation of tobacco mosaic virus movement protein binding to single-stranded nucleic acids. Plant Cell 4:397–411
    [Google Scholar]
  6. Gorbalenya A. E., Koonin E. V., Donchenko A. P., Blinov V. M. 1988; A conserved NTP-motif in putative helicases. Nature 333:22
    [Google Scholar]
  7. Gorbalenya A. E., Koonin E. V., Wolf Y. I. 1990; A new superfamily of putative NTP-binding domains encoded by genomes of small DNA and RNA viruses. FEBS Letters 262:145–148
    [Google Scholar]
  8. Hadaschik D., Klein M., Zimmermann H., Eggers H. J., Nelsen-Salz B. 1999; Dependence of echovirus-9 on the enteroviral replication inhibitor 2-(α-hydroxybenzyl)-benzimidazole (HBB) maps to the nonstructural protein 2C. Journal of Virology 73:10536–10539
    [Google Scholar]
  9. Kadaré G., Haenni A. L. 1997; Virus-encoded RNA helicases. Journal of Virology 71:2583–2590
    [Google Scholar]
  10. Klein M., Eggers H. J., Nelsen-Salz B. 1999; Echovirus-9 strain Barty nonstructural protein 2C has NTPase activity. Virus Research 65:155–160
    [Google Scholar]
  11. Klein M., Hadaschik D., Zimmermann H., Eggers H. J., Nelsen-Salz B. 2000; The picornavirus replication inhibitors HBB and guanidine in the echovirus-9 system: the significance of viral protein 2C. Journal of General Virology 81:895–901
    [Google Scholar]
  12. Lazinski D., Grzadzielska E., Das A. 1989; Sequence-specific recognition of RNA hairpins by bacteriophage antiterminators requires conserved arginine-rich motifs and is required for hepatitis delta virus RNA replication. Cell 59:207–218
    [Google Scholar]
  13. Mirzayan C., Wimmer E. 1992; Genetic analysis of an NTP-binding motif in poliovirus polypeptide 2C. Virology 189:547–555
    [Google Scholar]
  14. Mirzayan C., Wimmer E. 1994; Biochemical studies on poliovirus polypeptide 2C: evidence for ATPase activity. Virology 199:176–187
    [Google Scholar]
  15. Pincus S. E., Wimmer E. 1986; Production of guanidine-resistant and -dependent poliovirus mutants from cloned cDNA: mutations in polypeptide 2C are directly responsible for altered guanidine sensitivity. Journal of Virology 60:793–796
    [Google Scholar]
  16. Rodriguez P. L., Carrasco L. 1993; Poliovirus protein 2C has ATPase and GTPase activities. Journal of Biological Chemistry 268:8105–8110
    [Google Scholar]
  17. Rodriguez P. L., Carrasco L. 1995; Poliovirus protein 2C contains two regions involved in RNA binding activity. Journal of Biological Chemistry 270:10105–10112
    [Google Scholar]
  18. Tolskaya E. A., Romanova L. I., Kolesnikova M. S., Gmyl A. P., Gorbalenya A. P., Agol V. I. 1994; Genetic studies on the poliovirus 2C protein, an NTPase. A plausible mechanism of guanidine effect on the 2C function and evidence for the importance of 2C oligomerisation. Journal of Molecular Biology 236:1310–1323
    [Google Scholar]
  19. Zimmermann H., Eggers H. J., Nelsen-Salz B. 1996; Molecular cloning and sequence determination of the complete genome of the virulent echovirus-9 strain Barty. Virus Genes 12:149–154
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-81-10-2481
Loading
/content/journal/jgv/10.1099/0022-1317-81-10-2481
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