Immunological characterization of Toscana virus proteins

Immunological characterization of Toscana virus proteins The genome of Toscana virus ( Bunyaviridae family, Phlebovirus genus) consists of three single stranded RNA segments (L, M, S), with negative polarity. The L and M segments contain a single ORF in viral complementary sense and the S segment contains two ORFs in “ambisense” orientation. The M segment codes for three proteins in 3′–5′ genomic orientation: a 30 kDa non structural protein and two 65 kDa glycoproteins, G N , and G C . In this paper we report the expression in E. coli of the S segment ORFs and of three regions of the L ORF. The expressed proteins were used to produce monospecific polyclonal antibodies in mice. By using these antibodies the N and the NSs proteins were unequivocally assigned to the S viral-complementary and viral-sense ORFs, respectively, and the L protein to the L ORF. We have found that like N and L proteins, NSs protein is associated with the viral nucleocapsids in mature virions, suggesting its possible involvement in early events of viral replication. NSs protein was also found associated with cellular polysomes. In virus-infected cells the anti-L antibodies recognized proteins shorter than the full-length L protein, possibly products of L subgenomic segments. Interestingly these defective products were not found in mature virions, suggesting specific mechanisms in virion assembly. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Archives of Virology Springer Journals

Immunological characterization of Toscana virus proteins

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Publisher
Springer-Verlag
Copyright
Copyright © Wien by 1999 Springer-Verlag/
Subject
Legacy
ISSN
0304-8608
eISSN
1432-8798
D.O.I.
10.1007/s007050050717
Publisher site
See Article on Publisher Site

Abstract

The genome of Toscana virus ( Bunyaviridae family, Phlebovirus genus) consists of three single stranded RNA segments (L, M, S), with negative polarity. The L and M segments contain a single ORF in viral complementary sense and the S segment contains two ORFs in “ambisense” orientation. The M segment codes for three proteins in 3′–5′ genomic orientation: a 30 kDa non structural protein and two 65 kDa glycoproteins, G N , and G C . In this paper we report the expression in E. coli of the S segment ORFs and of three regions of the L ORF. The expressed proteins were used to produce monospecific polyclonal antibodies in mice. By using these antibodies the N and the NSs proteins were unequivocally assigned to the S viral-complementary and viral-sense ORFs, respectively, and the L protein to the L ORF. We have found that like N and L proteins, NSs protein is associated with the viral nucleocapsids in mature virions, suggesting its possible involvement in early events of viral replication. NSs protein was also found associated with cellular polysomes. In virus-infected cells the anti-L antibodies recognized proteins shorter than the full-length L protein, possibly products of L subgenomic segments. Interestingly these defective products were not found in mature virions, suggesting specific mechanisms in virion assembly.

Journal

Archives of VirologySpringer Journals

Published: Oct 1, 1999

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