1887

Abstract

Feline calicivirus (FCV) is a small positive-stranded RNA virus within the family . Its genome is 7690 nucleotides in length and encodes three open reading frames (ORFs). The smallest, ORF3, is located at the extreme 3′ end of the genome and can potentially encode a polypeptide of approximately 12 kDa. In this paper, we report the identification of an ORF3-encoded polypeptide in FCV-infected cells using an antiserum raised against a bacterially-expressed bacteriophage T7 gene 10-ORF3 fusion protein. Although a small mRNA of 0.5 kb, which could potentially encode ORF3, has been described, reports on the number and size of FCV subgenomic RNAs have varied considerably. To clarify the situation, RNAs from FCV-infected cells were labelled using [P]orthophosphate, an approach which provided definitive data. Only two RNA species were detected, the genomic RNA and a subgenomic mRNA of 2.4 kb. The 5′ end of the subgenomic mRNA was mapped to position 5227 on the genomic RNA using RNA sequencing and primer extension methods. RNA isolated from FCV-infected cells in which no subgenomic RNA smaller than 2.4 kb was detectable directed the synthesis in rabbit reticulocyte lysate of the ORF3-encoded polypeptide. Furthermore, a synthetic RNA copy of the 2.4 kb subgenomic mRNA of FCV, containing both ORF2 and 3, directed the synthesis of both ORF2 and ORF3 polypeptides in the translation system. These data strongly suggest that ORF3 is expressed from the 2.4 kb subgenomic RNA and that this RNA is functionally bicistronic. The possible mechanisms by which ORF3 is expressed are discussed.

Loading

Article metrics loading...

/content/journal/jgv/10.1099/0022-1317-77-1-123
1996-01-01
2022-01-21
Loading full text...

Full text loading...

/deliver/fulltext/jgv/77/1/JV0770010123.html?itemId=/content/journal/jgv/10.1099/0022-1317-77-1-123&mimeType=html&fmt=ahah

References

  1. Black D. N., Burroughs J. N. B., Harris T. J. R., Brown F. 1978; The structure and replication of calicivirus RNA. Nature 274:614–615
    [Google Scholar]
  2. Brierley I. 1995; Ribosomal frameshifting on viral RNAs. Journal of General Virology 76:1885–1892
    [Google Scholar]
  3. Carter M. J. 1990; Transcription of feline calicivirus RNA. Archives of Virology 114:143–152
    [Google Scholar]
  4. Carter M. J., Milton I. D., Meanger J., Bennett M., Gaskell R. M., Turner P. C. 1992a; The complete nucleotide sequence of feline calicivirus. Virology 190:443–448
    [Google Scholar]
  5. Carter M. J., Milton I. D., Turner P. C., Meanger J., Bennett M., Gaskell R. M. 1992b; Identification and sequence determination of the capsid gene of feline calicivirus. Archives of Virology 122:223–235
    [Google Scholar]
  6. Ehresmann D. W., Schaffer F. L. 1977; RNA synthesized in calicivirus infected cells is atypical of picornaviruses. Journal of Virology 22:572–576
    [Google Scholar]
  7. Ehresmann D. W., Schaffer F. L. 1979; Calicivirus intracellular RNA: fractionation of 18-22S RNA and lack of typical 5′-methylated caps on 36S and 22S San Miguel sea lion virus RNA. Virology 95:251–255
    [Google Scholar]
  8. Horvath C. M., Williams N. A., Lamb R. A. 1990; Eukaryotic coupled translation of tandem cistrons: identification of the influenza B virus BM2 polypeptide. EMBO Journal 9:2639–2647
    [Google Scholar]
  9. Jiang X., Wang M., Wang K., Estes M. K. 1993; Sequence and genomic organization of Norwalk virus. Virology 195:51–61
    [Google Scholar]
  10. Kozak M. 1987; Effects of intercistronic distance on the efficiency of reinitiation by eukaryotic ribosomes. Molecular and Cellular Biology 7:3438–3445
    [Google Scholar]
  11. Kozak M. 1989; The scanning model for translation: an update. Journal of Cell Biology 108:229–241
    [Google Scholar]
  12. Liu D. X., Inglis S. C. 1992; Internal entry of ribosomes on a tricistronic mRNA encoded by IBV. Journal of Virology 66:6143–6154
    [Google Scholar]
  13. Meyers G., Wirblich C., Thiel H. 1991; Rabbit hemorrhagic disease virus - molecular cloning and nucleotide sequencing of a calicivirus genome. Virology 184:664–676
    [Google Scholar]
  14. Milton I. D. M., Vlasak R., Nowotny N., Rodak L., Carter M. J. 1992; Genomic 3′ terminal sequence comparison of three isolates of rabbit haemorrhagic disease virus. FEMS Microbiology Letters 93:37–42
    [Google Scholar]
  15. Neill J. D. 1990; Nucleotide sequence of a region of the feline calicivirus genome which encodes picornavirus-like RNA-dependent RNA polymerase, cysteine protease and 2C polypeptides. Virus Research 17:145–160
    [Google Scholar]
  16. Neill J. D. 1992; Nucleotide sequence of the capsid protein gene of two serotypes of San Miguel sea lion virus: identification of conserved and non-conserved amino acid sequences among calicivirus capsid proteins. Virus Research 24:211–222
    [Google Scholar]
  17. Neill J. D., Mengeling W. L. 1988; Further characterisation of the virus-specific RNAs in feline calicivirus infected cells. Virus Research 11:59–72
    [Google Scholar]
  18. Neill J. D., Reardon I. M., Heinrickson R. L. 1991; Nucleotide sequence and expression of the capsid protein gene of feline calicivirus. Journal of Virology 65:5440–5447
    [Google Scholar]
  19. Oh S., Sarnow P. 1993; Gene regulation: translational initiation by internal ribosome binding. Current Opinions in Genetics and Development 3:295–300
    [Google Scholar]
  20. Peabody D. S., Berg P. 1986; Termination-reinitiation occurs in the translations of mammalian cell mRNAs. Molecular and Cellular Biology 6:2695–2703
    [Google Scholar]
  21. Seal B. S., Ridpath J. F., Mengeling W. L. 1993; Analysis of feline calicivirus capsid protein genes: identification of variable antigenic determinant regions of the protein. Journal of General Virology 74:2519–2524
    [Google Scholar]
http://instance.metastore.ingenta.com/content/journal/jgv/10.1099/0022-1317-77-1-123
Loading
/content/journal/jgv/10.1099/0022-1317-77-1-123
Loading

Data & Media loading...

Most cited this month Most Cited RSS feed

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