1887

Abstract

Among randomly mutagenized clones derived from an infectious cDNA copy of genomic RNA of (ASGV), we previously identified a clone, pRM21, whose transcript (ASGV-RM21) does not induce any symptoms characteristic of the original (wild-type) cDNA clone (ASGV-wt) in several host plants. Interestingly, ASGV-RM21 has only a single, translationally silent nucleotide substitution, U to C, at nucleotide 4646 of the viral genome within open reading frame (ORF) 1. Here, we characterize and verify this unprecedented silent-mutation-induced attenuation of symptoms in infected plants. Northern and Western blot analyses showed that less ASGV-RM21 accumulates in host plants than ASGV-wt. In addition, two more silent substitutions, U to A and U to G, constructed by site-directed mutagenesis at the same nucleotide (4646), also induced attenuated symptoms. This is the first report that a single silent substitution attenuates virus-infection symptoms and implicates a novel determinant of disease symptom severity.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/vir.0.19179-0
2003-09-01
2024-03-28
Loading full text...

Full text loading...

/deliver/fulltext/jgv/84/9/vir842579.html?itemId=/content/journal/jgv/10.1099/vir.0.19179-0&mimeType=html&fmt=ahah

References

  1. Andersen K., Johansen I. E. 1998; A single conserved amino acid in the coat protein gene of pea seed-borne mosaic potyvirus modulates the ability of the virus to move systemically in Chenopodium quinoa . Virology 241:304–311
    [Google Scholar]
  2. Banerjee N., Wang J.-Y., Zaitlin M. 1995; A single nucleotide change in the coat protein gene of tobacco mosaic virus is involved in the induction of severe chlorosis. Virology 207:234–239
    [Google Scholar]
  3. de Assis Filho F. M., Paguio O. R., Sherwood J. L., Deom C. M. 2002; Symptom induction by Cowpea chlorotic mottle virus on Vigna unguiculata is determined by amino acid residue 151 in the coat protein. J Gen Virol 83:879–883
    [Google Scholar]
  4. Fujita Y., Mise K., Okuno T., Ahlquist P., Furusawa I. 1996; A single codon change in a conserved motif of a Bromovirus movement protein gene confers compatibility with a new host. Virology 223:283–291
    [Google Scholar]
  5. Greener A., Callahan M., Jerpseth B. 1997; An efficient random mutagenesis technique using an E. coli mutator strain. Mol Biotechnol 7:189–195
    [Google Scholar]
  6. Hagiwara K., Ichiki T. U., Ogawa Y., Omura T., Tsuda S. 2002; A single amino acid substitution in 126-kDa protein of Pepper mild mottle virus associates with symptom attenuation in pepper; the complete nucleotide sequence of an attenuated strain, C-1421. Arch Virol 147:833–340
    [Google Scholar]
  7. Holt C. A., Hodgson R. A. J., Coker F. A., Beachy R. N., Nelson R. S. 1990; Characterization of the masked strain of tobacco mosaic virus: identification of the region responsible for symptom attenuation by analysis of an infectious cDNA clone. Mol Plant Microbe Interact 3:417–423
    [Google Scholar]
  8. Lewandowski D. J., Dawson W. O. 1993; A single amino acid change in tobacco mosaic virus replicase prevents symptom production. Mol Plant Microbe Interact 6:157–160
    [Google Scholar]
  9. Lu X., Hirata H., Yamaji Y., Ugaki M., Namaba S. 2001; Random mutagenesis in a plant viral genome using a DNA repair-deficient mutator Escherichia coli strain. J Virol Methods 94:37–43
    [Google Scholar]
  10. Magome H., Terauchi H., Yoshikawa N., Takahashi T. 1997; Analysis of double-stranded RNA in tissues infected with apple stem grooving capillovirus. Ann Phytopathol Soc Jpn 63:450–454
    [Google Scholar]
  11. Moreno I. M., Bernal J. J., de Blas B. G., Rodriguez-Cerezo E., Gracıa-Arenal F. 1997; The expression level of the 3a movement protein determines differences in severity of symptoms between two strains of tomato aspermy cucumovirus. Mol Plant Microbe Interact 10:171–179
    [Google Scholar]
  12. Nishiguchi M., Kikuchi S., Kiho Y., Ohno T., Meshi T., Okada Y. 1985; Molecular basis of plant viral virulence; the complete nucleotide sequence of an attenuated strain of tobacco mosaic virus. Nucleic Acids Res 13:5585–5590
    [Google Scholar]
  13. Nishio T., Kawai A., Takahashi T., Namba S., Yamashita S. 1989; Purification and properties of citrus tatter leaf virus. Ann Phytopathol Soc Jpn 55:254–258
    [Google Scholar]
  14. Ohira K., Ito T., Kawai A., Namba S., Kusumi T., Tsuchizaki T. 1994; Nucleotide sequence of the 3′-terminal region of citrus tatter leaf virus RNA. Virus Genes 8:169–172
    [Google Scholar]
  15. Ohira K., Namba S., Rozanov M., Kusumi T., Tsuchizaki T. 1995; Complete sequence of an infectious full-length cDNA clone of citrus tatter leaf capillovirus: comparative sequence analysis of capillovirus genomes. J Gen Virol 76:2305–2309
    [Google Scholar]
  16. Rao A. L. N., Grantham G. L. 1995; A spontaneous mutation in the movement protein gene of brome mosaic virus modulates symptom phenotype in Nicotiana benthamiana . J Gen Virol 69:2689–2691
    [Google Scholar]
  17. Rodrıguez-Cerezo E., Klein P. G., Shaw J. G. 1991; A determinant of disease symptom severity is located in the 3′-terminal noncoding region of the RNA of a plant virus. Proc Natl Acad Sci U S A 88:9863–9867
    [Google Scholar]
  18. Sambrook J., Russell D. W. 2001a Molecular Cloning: a Laboratory Manual , 3rd edn. vol 3 pp. A8.40–A8.55 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  19. Sambrook J., Russell D. W. 2001b Molecular Cloning: a Laboratory Manual , 3rd edn. vol 1 pp. 7.31–7.50 Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  20. Shintaku M. H., Palukaitis P. 1992; A single amino acid substitution in the coat protein of cucumber mosaic virus induces chlorosis in tobacco. Plant Cell 4:751–757
    [Google Scholar]
  21. Szilassy D., Salánki K., Balázs E. 1999; Stunting induced by cucumber mosaic cucumovirus-infected Nicotiana glutinosa is determined by a single amino acid residue in the coat protein. Mol Plant Microbe Interact 12:1105–1113
    [Google Scholar]
  22. Takeshita M., Suzuki M., Takanami Y. 2001; Combination of amino acids in the 3a protein and the coat protein of Cucumber mosaic virus determines symptom expression and viral spread in bottle gourd. Arch Virol 146:697–711
    [Google Scholar]
  23. van der Vossen E. A. G., Neeleman L., Bol J. F. 1996; The 5′ terminal sequence of alfalfa mosaic virus RNA3 is dispensable for replication and contains a determinant for symptom formation. Virology 221:271–280
    [Google Scholar]
  24. Verwoerd T. C., Dekker B. M., Hoekema A. 1989; A small-scale procedure for the rapid isolation of plant RNAs. Nucleic Acids Res 17:2362
    [Google Scholar]
  25. Yoshikawa N., Takahashi T. 1992; Evidence for translation of apple stem grooving capillovirus genomic RNA. J Gen Virol 73:1313–1315
    [Google Scholar]
  26. Yoshikawa N., Sasaki E., Kato M., Takahashi T. 1992; The nucleotide sequence of apple stem grooving capillovirus genome. Virology 191:98–105
    [Google Scholar]
  27. Yoshikawa N., Imaizumi M., Takahashi T., Inouye N. 1993; Striking similarities between the nucleotide sequence and genome organization of citrus tatter leaf and apple stem grooving capilloviruses. J Gen Virol 74:2743–2747
    [Google Scholar]
  28. Zhang L., Hanada K., Palukaitis P. 1994; Mapping local and systemic symptom determinants of cucumber mosaic cucumovirus in tobacco. J Gen Virol 75:3185–3191
    [Google Scholar]
  29. Zuker M., Jacobson A. B. 1998; Using reliability information to annotate RNA secondary structures. RNA 4:669–679
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/vir.0.19179-0
Loading
/content/journal/jgv/10.1099/vir.0.19179-0
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