Mutation of cysteine 171 of pestivirus E rns RNase prevents homodimer formation and leads to attenuation of classical swine fever virus.
Pestiviruses represent important pathogens of farm animals that have evolved unique strategies and functions to stay within their host populations. E(rns), a structural glycoprotein of pestiviruses, exhibits RNase activity and represents a virulence factor of the viruses. E(rns) forms disulfide linked homodimers that are found in virions and virus-infected cells. Mutation or deletion of cysteine 171, the residue engaged in intermolecular disulfide bond formation, results in loss of dimerization as tested in coprecipitation and native protein gel electrophoresis analyses. Nevertheless, stable virus mutants with changes affecting cysteine codon 171 could be recovered in tissue culture. These mutants grew almost as well as the parental viruses and exhibited an RNase-positive phenotype. E(rns) dimerization-negative mutants of classical swine fever virus were found to be attenuated in pigs even though the virus clearly replicated and induced a significant neutralizing antibody response in the animals.
Tews
Birke Andrea
BA
Institut für Immunologie, Friedrich-Loeffler-Institut, Paul-Ehrlich-Str. 28, D-72076 Tübingen, Germany.
Schürmann
Eva-Maria
EM
Meyers
Gregor
G
D000818:Animals; D000914:Antibodies, Viral / Q000097:blood; D002460:Cell Line; D006691:Classical Swine Fever / Q000276:immunology / Q000821:virology; D006692:Classical Swine Fever Virus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D003545:Cysteine / Q000235:genetics*; D007370:Interferon Type I / Q000097:blood; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D009154:Mutation; D010957:Plasmids; D055503:Protein Multimerization; D012260:Ribonucleases / Q000378:metabolism*; D013552:Swine / Q000276:immunology / Q000821:virology; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism
D000914:Antibodies, Viral; C466501:E(rns) protein, classic swine fever virus; D007370:Interferon Type I; D008562:Membrane Glycoproteins; D014759:Viral Envelope Proteins; D012260:Ribonucleases; D003545:Cysteine