Replication and amplification of novel vesicular stomatitis virus minigenomes encoding viral structural proteins

E. A. Stillman, J. K. Rose, Michael Whitt

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

We have developed a system in which vesicular stomatitis virus (VSV) minigenomes encoding viral structural proteins can be expressed from plasmids. These RNAs can be replicated, transcribed, and packaged into infectious particles when coexpressed with the other VSV proteins. The minigenomes contain either the glycoprotein (G protein) gene (GMG [stands for G minigenome]) or both the G and matrix (M) protein genes (GMMG [stands for G/M minigenome]) from the Indiana serotype of VSV flanked by the trailer and leader regions from the wild-type VSV genome. Northern (RNA) blot analysis showed that the minigenome RNAs were replicated and that a positive-sense replicative intermediate was synthesized when coexpressed with the nucleocapsid (N) protein and the two VSV polymerase proteins (phosphoprotein [P] and the large catalytic subunit [L]) in vivo. In addition, functional mRNAs were transcribed from the minigenome templates, and the appropriate encoded proteins were expressed. Expression of the G and M proteins from GMMG resulted in the assembly and release of infectious particles that could be passaged on cells expressing the N, P, and L proteins only. Amplification occurred during successive passages, and after four passages approximately 30% of the cells expressed both the G and M proteins. Analysis of the RNAs produced in the GMMG-infected cells also showed that the minigenomes accurately reproduced all of the replicative and transcriptional events that normally occur in a VSV-infected cell. GMMG is therefore a novel type of defective particle which encodes functional viral proteins critical to its own propagation.

Original languageEnglish (US)
Pages (from-to)2946-2953
Number of pages8
JournalJournal of Virology
Volume69
Issue number5
StatePublished - 1995

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Viral Structural Proteins
Vesiculovirus
Vesicular Stomatitis
viral proteins
structural proteins
Viruses
Proteins
RNA
proteins
Vesicular stomatitis Indiana virus
Nucleocapsid Proteins
cells
Phosphoproteins
nucleocapsid
trailers
phosphoproteins
Viral Proteins
Northern Blotting
protein subunits
Catalytic Domain

All Science Journal Classification (ASJC) codes

  • Immunology

Cite this

Replication and amplification of novel vesicular stomatitis virus minigenomes encoding viral structural proteins. / Stillman, E. A.; Rose, J. K.; Whitt, Michael.

In: Journal of Virology, Vol. 69, No. 5, 1995, p. 2946-2953.

Research output: Contribution to journalArticle

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