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Journal of Virology, May 2009, p. 4823-4834, Vol. 83, No. 10
0022-538X/09/$08.00+0     doi:10.1128/JVI.01710-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Mutation of Cysteine 171 of Pestivirus Erns RNase Prevents Homodimer Formation and Leads to Attenuation of Classical Swine Fever Virus{triangledown}

Birke Andrea Tews,{dagger} Eva-Maria Schürmann, and Gregor Meyers*

Institut für Immunologie, Friedrich-Loeffler-Institut, D-72001 Tübingen, Germany

Received 12 August 2008/ Accepted 20 February 2009

Pestiviruses represent important pathogens of farm animals that have evolved unique strategies and functions to stay within their host populations. Erns, a structural glycoprotein of pestiviruses, exhibits RNase activity and represents a virulence factor of the viruses. Erns 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. Erns 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.

* Corresponding author. Mailing address: Institut für Immunologie, Friedrich-Loeffler-Institut, Paul-Ehrlich-Str. 28, D-72076 Tübingen, Germany. Phone: 49 7071-9670. Fax: 49 7071-967303. E-mail: gregor.meyers{at}fli.bund.de

{triangledown} Published ahead of print on 4 March 2009.

{dagger} Present address: Institut de Biologie de Lille (UMR8161), Institut Pasteur de Lille, Lille, France.

Journal of Virology, May 2009, p. 4823-4834, Vol. 83, No. 10
0022-538X/09/$08.00+0     doi:10.1128/JVI.01710-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





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