Biochemical characterization of salmon pancreas disease virus

• Published in: Journal of general virology 2000-03, Vol.81 (3), p.813-820
• Main Authors: Welsh, Michael, Weston, Jonathan, Borghmans, Borghert Jan, Mackie, Dermot, Rowley, Helen, Nelson, Robert, McLoughlin, Marian, Todd, Daniel
• Format: Article
• Language: English
• Subjects: Alphavirus; Alphavirus - chemistry; Alphavirus - genetics; Alphavirus - isolation & purification; Animals; Antibodies, Monoclonal; Antibodies, Viral; Base Sequence; DNA Primers - genetics; double prime E1 protein; double prime E2 protein; fish culture; Fish Diseases - virology; Marine; Mice; Oncorhynchus tshawytscha; Pancreatic Diseases - veterinary; Pancreatic Diseases - virology; RNA, Viral - chemistry; RNA, Viral - genetics; Salmo salar; Salmon pancreas disease virus; Togaviridae; Viral Proteins - chemistry
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-81-3-813

Department of Veterinary Sciences, The Queen’s University of Belfast, Stormont, Belfast BT4 3SD, UK 1 Veterinary Sciences Division, Department of Agriculture for Northern Ireland, Stormont, Belfast BT4 3SD, UK 2 Author for correspondence: Michael Welsh. Present address: Department of Bacteriology, Veterinary Sciences Division, Department of Agriculture for Northern Ireland, Stormont, Belfast BT4 3SD, UK. Fax [phone]. e-mail michael.welsh{at}dani.gov.uk Salmon pancreas disease virus (SPDV) has been shown to cause severe economic losses in farmed Atlantic salmon ( Salmo salar ) and has been reported to occur in Europe, Scandinavia and the United States. This paper describes the biochemical characterization of SPDV in terms of its RNA and protein composition. SPDV was purified by precipitation from infected Chinook salmon embryo (CHSE-214) cell-culture supernatant and sucrose density-gradient centrifugation. Fractions containing virus were identified by an immunodot blot assay using an SPDV-specific MAb. Two major proteins with molecular masses of approximately 55 and 50 kDa, putatively identified as the E1 and E2 alphavirus glycoproteins respectively, were detected when purified virus preparations were analysed by PAGE. Radiolabelling experiments indicated that SPDV infection of CHSE-214 cells did not shut-off host-cell protein synthesis, making attempts to identify virus-specific proteins unsuccessful. However, radioimmunoprecipitation assay (RIPA) experiments showed that two SPDV-specific MAbs reacted with a protein in the 50–55 kDa range. Northern blot hybridization with cloned cDNA probes indicated that infected cells contained RNA species of approximately 11·4 and 4 kb, which correspond to the genomic and subgenomic RNAs specified by SPDV. The results described are consistent with SPDV being characterized as an alphavirus.