1887

Abstract

To study the proteolytic processing of the potato leafroll virus replicase proteins, the multidomain P1 protein with a c- epitope tag attached at the N terminus was expressed in insect cells by using the baculovirus system. Western blotting showed that P1 was cleaved at a site upstream of the serine protease domain, in addition to the cleavage site downstream of the protease domain. Mutational analysis showed that the serine protease domain within P1 was responsible for this cleavage. To characterize this novel cleavage site further, a portion of the P1 protein comprising the protease domain and the two cleavage sites was expressed in . A similar cleavage event was observed in bacteria and was abolished when the P1 protease was inactivated by mutation. Peptide-sequencing studies indicated that this cleavage occurred at a Glu/Arg junction, separating the N-terminal 204 residues from the serine protease domain of P1.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.82627-0
2007-05-01
2024-04-16
Loading full text...

Full text loading...

/deliver/fulltext/jgv/88/5/1620.html?itemId=/content/journal/jgv/10.1099/vir.0.82627-0&mimeType=html&fmt=ahah

References

  1. Ashoub A., Rohde W., Prüfer D. 1998; In planta transcription of a second subgenomic RNA increases the complexity of the subgroup 2 luteovirus genome. Nucleic Acids Res 26:420–426 [CrossRef]
    [Google Scholar]
  2. Dougherty W. G., Semler B. L. 1993; Expression of virus-encoded proteinases: functional and structural similarities with cellular enzymes. Microbiol Rev 57:781–822
    [Google Scholar]
  3. Godeny E. K., Chen L., Kumar S. N., Methven S. L., Koonin E. V., Brinton M. A. 1993; Complete genomic sequence and phylogenetic analysis of the lactate dehydrogenase-elevating virus (LDV). Virology 194:585–596 [CrossRef]
    [Google Scholar]
  4. Harrison B. D. 1984 Potato leafroll virus (CMI/AAB Description of Plant Viruses no 291 Kew, UK: CMI/AAB;
    [Google Scholar]
  5. Jaag H. M., Kawchuk L., Rohde W., Fischer R., Emans N., Prüfer D. 2003; An unusual internal ribosomal entry site of inverted symmetry directs expression of a potato leafroll polerovirus replication-associated protein. Proc Natl Acad Sci U S A 100:8939–8944 [CrossRef]
    [Google Scholar]
  6. Keese P., Martin R. R., Kawchuk L. M., Waterhouse P. M., Gerlach W. L. 1990; Nucleotide sequences of an Australian and a Canadian isolate of potato leafroll luteovirus and their relationships with two European isolates. J Gen Virol 71:719–724 [CrossRef]
    [Google Scholar]
  7. Koonin E. V., Dolja V. V. 1993; Evolution and taxonomy of positive-strand RNA viruses: implications of comparative analysis of amino acid sequences. Crit Rev Biochem Mol Biol 28:375–430 [CrossRef]
    [Google Scholar]
  8. Li X., Ryan M. D., Lamb J. W. 2000; Potato leafroll virus protein P1 contains a serine proteinase domain. J Gen Virol 81:1857–1864
    [Google Scholar]
  9. Mayo M. A., Ziegler-Graff V. 1996; Molecular biology of luteoviruses. Adv Virus Res 46:413–460
    [Google Scholar]
  10. Mayo M. A., Robinson D. J., Jolly C. A., Hyman L. 1989; Nucleotide sequence of potato leafroll luteovirus RNA. J Gen Virol 70:1037–1051 [CrossRef]
    [Google Scholar]
  11. Miller J. S., Mayo M. A. 1991; The location of the 5′ end of the potato leafroll luteovirus subgenomic coat protein mRNA. J Gen Virol 72:2633–2638 [CrossRef]
    [Google Scholar]
  12. Miller W. A., Dinesh-Kumar S. P., Paul C. P. 1995; Luteovirus gene expression. Crit Rev Plant Sci 14:179–211 [CrossRef]
    [Google Scholar]
  13. Pringle C. R. 1998; Virus taxonomy – San Diego 1998. Arch Virol 143:1449–1459 [CrossRef]
    [Google Scholar]
  14. Prüfer D., Tacke E., Schmitz J., Kull B., Kaufmann A., Rohde W. 1992; Ribosomal frameshifting in plants: a novel signal directs the –1 frameshift in the expression of the putative viral replicase of potato leafroll luteovirus. EMBO J 11:1111–1117
    [Google Scholar]
  15. Prüfer D., Kawchuk L., Monecke M., Nowok S., Fischer R., Rohde W. 1999; Immunological analysis of potato leaf roll luteovirus (PLRV) P1 expression identifies a 25 kD RNA-binding protein derived via P1 processing. Nucleic Acids Res 27:421–425 [CrossRef]
    [Google Scholar]
  16. Reutenauer A., Ziegler-Graff V., Lot H., Scheidecker D., Guilley H., Richards K., Jonard G. 1993; Identification of beet western yellows luteovirus genes implicated in viral replication and particle morphogenesis. Virology 195:692–699 [CrossRef]
    [Google Scholar]
  17. Sadowy E., Juszczuk M., David C., Gronenborn B., Hulanicka M. D. 2001a; Mutational analysis of the proteinase function of Potato leafroll virus . J Gen Virol 82:1517–1527
    [Google Scholar]
  18. Sadowy E., Maasen A., Juszczuk M., David C., Zagorski-Ostoja W., Gronenborn B., Hulanicka M. D. 2001b; The ORF0 product of Potato leafroll virus is indispensable for virus accumulation. J Gen Virol 82:1529–1532
    [Google Scholar]
  19. Snijder E. J., Wassenaar A. L., Spaan W. J. 1994; Proteolytic processing of the replicase ORF1a protein of equine arteritis virus. J Virol 68:5755–5764
    [Google Scholar]
  20. Snijder E. J., Wassenaar A. L., van Dinten L. C., Spaan W. J., Gorbalenya A. E. 1996; The arterivirus nsp4 protease is the prototype of a novel group of chymotrypsin-like enzymes, the 3C-like serine proteases. J Biol Chem 271:4864–4871 [CrossRef]
    [Google Scholar]
  21. Tacke E., Prüfer D., Salamini F., Rohde W. 1990; Characterization of a potato leafroll luteovirus subgenomic RNA: differential expression by internal translation initiation and UAG suppression. J Gen Virol 71:2265–2272 [CrossRef]
    [Google Scholar]
  22. van der Wilk F., Huisman M. J., Cornelissen B. J., Huttinga H., Goldbach R. 1989; Nucleotide sequence and organization of potato leafroll virus genomic RNA. FEBS Lett 245:51–56 [CrossRef]
    [Google Scholar]
  23. van der Wilk F., Verbeek M., Dullemans A. M., van den Heuvel J. F. 1997; The genome linked protein of potato leafroll virus is located downstream of the putative protease domain of the ORF1 product. Virology 234:300–303 [CrossRef]
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.82627-0
Loading
/content/journal/jgv/10.1099/vir.0.82627-0
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