Detection of the ORF3 polypeptide of feline calicivirus in infected cells and evidence for its expression from a single, functionally bicistronic, subgenomic mRNA

Herbert, Terence and Brierley, I. and Brown, T.D.K. (1996) Detection of the ORF3 polypeptide of feline calicivirus in infected cells and evidence for its expression from a single, functionally bicistronic, subgenomic mRNA. Journal of General Virology, 77 (1). pp. 123-127. ISSN 0022-1317

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Abstract

Feline calicivirus (FCV) is a small positive-stranded RNA virus within the family Caliciviridae. 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 in vivo using [32P]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 in vitro 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.

Additional Information:cited By 61 The final published version of this article can be accessed online at http://jgv.microbiologyresearch.org/content/journal/jgv/10.1099/0022-1317-77-1-123
Keywords:ORF3 polypeptide, feline calicivirus, mRNA
Subjects:B Subjects allied to Medicine > B200 Pharmacology, Toxicology and Pharmacy
Divisions:College of Science > School of Pharmacy
ID Code:28242
Deposited On:13 Sep 2018 13:49

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