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Journal of General Virology (2000), 81, 939-947.
© 2000 Society for General Microbiology

Animal: RNA Viruses

A short leucine-rich sequence in the Borna disease virus p10 protein mediates association with the viral phospho- and nucleoproteins

Thorsten Wolff1, Rene Pfleger1, Tatjana Wehner2, Jens Reinhardt1 and Juergen A. Richt2

Institut für Virologie, Philipps-Universität Marburg, Robert-Koch-Str. 17, 35037 Marburg, Germany1
Institut für Virologie, Justus Liebig Universität Giessen, Frankfurter Str. 107, 35392 Giessen, Germany2

Author for correspondence: Thorsten Wolff. Present address: Robert-Koch-Institut, Nordufer 20, 13353 Berlin, Germany. Fax +49 30 4547 2328. e-mail wolfft{at}rki.de

Borna disease virus (BDV) is unique among the non-segmented negative-strand RNA viruses of animals and man because it transcribes and replicates its genome in the nucleus of the infected cell. It has recently been discovered that BDV expresses a gene product of 87 amino acids, the p10 protein, from an open reading frame that overlaps with the gene encoding the viral p24 phosphoprotein. In addition, the p10 protein has been localized to intranuclear BDV-specific clusters containing viral antigens. Here, characterization of p10 interactions with the viral nucleoprotein p38/p39 and the p24 phosphoprotein is reported. Immunoaffinity chromatography demonstrated the presence of high-salt stable complexes of p10 containing the p24 and p38/p39 proteins in extracts of BDV-infected cells. Analyses in the yeast two-hybrid system and biochemical co-precipitation experiments suggested that the p10 protein binds directly to the p24 phosphoprotein and indirectly to the viral nucleoprotein. Mutational analysis demonstrated that a leucine-rich stretch of amino acids at positions 8–15 within the p10 protein is critical for interaction with p24. Furthermore, binding of p10 to the viral phosphoprotein was shown to be important for association with the BDV-specific intranuclear clusters that may represent the sites of virus replication and transcription in infected cells. These findings are discussed with respect to possible roles for the p10 protein in viral RNA synthesis or ribonucleoprotein transport.




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