Reovirus growth in cell culture does not require the full complement of viral proteins: Identification of a σ1s-null mutant

Steven Rodgers, Jodi L. Connolly, James D. Chappell, Terence S. Dermody

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

The reovirus σ1s protein is a 14-kDa nonstructural protein encoded by the S1 gene segment. The S1 gene has been linked to many properties of reovirus, including virulence and induction of apoptosis. Although the function of σ1s is not known, the σ1s open reading frame is conserved in all S1 gene sequences determined to date. In this study, we identified and characterized a variant of type 3 reovirus, T3C84-MA, which does not express σ1s. To facilitate these experiments, we generated two monoclonal antibodies (MAbs) that bind different epitopes of the σ1s protein. Using these MAbs in immunoblot and immunofluorescence assays, we found that L929 (L) cells infected with T3C84-MA do not contain σ1s. To determine whether σ1s is required for reovirus infection of cultured cells, we compared the growth of T3C84-MA and its parental strain, T3C84, in L cells and Madin-Darby canine kidney (MDCK) cells. After 48 h of growth, yields of T3C84-MA were equivalent to yields of T3C84 in L cells and were fivefold lower than yields of T3C84 in MDCK cells. After 7 days of growth following adsorption at a low multiplicity of infection, yields of T3C84-MA and T3C84 did not differ significantly in either L cells or MDCK cells. To determine whether σ1s is required for apoptosis induced by reovirus infection, T3C84-MA and T3C84 were tested for their capacity to induce apoptosis, using an acridine orange staining assay. In these experiments, the percentages of apoptotic cells following infection with T3C84-MA and T3C84 were equivalent. These findings indicate that nonstructural protein σ1s is not required for reovirus growth in cell culture and does not influence the capacity of reovirus to induce apoptosis. Therefore, reovirus replication does not require the full complement of virally encoded proteins.

Original languageEnglish
Pages (from-to)8597-8604
Number of pages8
JournalJournal of Virology
Volume72
Issue number11
StatePublished - Nov 1 1998
Externally publishedYes

Fingerprint

Reoviridae
viral proteins
Viral Proteins
Complement System Proteins
complement
cell culture
Cell Culture Techniques
Madin Darby Canine Kidney Cells
Reoviridae Infections
mutants
Apoptosis
Growth
Proteins
kidney cells
apoptosis
Mammalian orthoreovirus 3
Monoclonal Antibodies
Genes
Acridine Orange
proteins

ASJC Scopus subject areas

  • Immunology

Cite this

Reovirus growth in cell culture does not require the full complement of viral proteins : Identification of a σ1s-null mutant. / Rodgers, Steven; Connolly, Jodi L.; Chappell, James D.; Dermody, Terence S.

In: Journal of Virology, Vol. 72, No. 11, 01.11.1998, p. 8597-8604.

Research output: Contribution to journalArticle

Rodgers, Steven ; Connolly, Jodi L. ; Chappell, James D. ; Dermody, Terence S. / Reovirus growth in cell culture does not require the full complement of viral proteins : Identification of a σ1s-null mutant. In: Journal of Virology. 1998 ; Vol. 72, No. 11. pp. 8597-8604.
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