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Publication type: Journal Article
Document type: Full Paper

Year: 1999

Author(s): Meyers, G; Saalmüller, A; Büttner, M

Title: Mutations abrogating the RNase activity in glycoprotein E(rns) of the pestivirus classical swine fever virus lead to virus attenuation.

Source: J Virol (73), 12 10224-10235.


Authors Vetmeduni Vienna:

Saalmüller Armin


Abstract:
Classical swine fever (CSF) is a severe hemorrhagic disease of swine caused by the pestivirus CSF virus (CSFV), Amino acid exchanges or deletions introduced by site-directed mutagenesis into the putative active site of the RNase residing in the glycoprotein E-rns CSFV abolished the enzymatic activity of this protein, as demonstrated with an RNase test suitable for detection of the enzymatic activity in crude cell extracts, Incorporation of the altered sequences into an infectious CSFV clone resulted in recovery of viable viruses upon RNA transfection, except for a variant displaying a deletion of the histidine codon at position 297 of the long open reading frame. These RNase-negative virus mutants displayed growth characteristics in tissue culture that were undistinguishable from wild-type virus and were stable for at least seven passages. In contrast to animals inoculated with an RNase-positive control virus, infection of piglets with an RNase-negative mutant containing a deletion of the histidine codon 346 of the open reading frame did not lead to CSF. Neither fever nor extended viremia could be detected. Animals infected with this mutant did not show decrease of peripheral B cells, a characteristic feature of CSF in swine. Animal experiments with four other mutants with either exchanges of codons 297 or 346 or double exchanges of both codons 297 and 346 showed that all these RNase-negative mutants were attenuated. All viruses with mutations affecting codon 346 were completely apathogenic, whereas those containing only changes of codon 297 consistently induced clinical symptoms for several days, followed by sudden recovery. Analyses of reisolated viruses gave no indication for the presence of revertants in the infected animals.

Keywords Pubmed: Amino Acid Sequence - : Animals - : Cell Line - : Classical swine fever virus - enzymology : Cricetinae - enzymology : Gene Deletion - enzymology : Glycoproteins - genetics : Molecular Sequence Data - genetics : Point Mutation - genetics : Ribonucleases - genetics : Swine - genetics : Viral Envelope Proteins - genetics

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