Cells that express all five proteins of vesicular stomatitis virus from cloned cDNAs support replication, assembly, and budding of defective interfering particles

Asit K. Pattnaik, Gail W. Wertz

Research output: Contribution to journalArticle

55 Scopus citations

Abstract

An alternative approach to structure-function analysis of vesicular stomatitis virus (VSV) gene products and their interactions with one another during each phase of the viral life cycle is described. We showed previously by using the vaccinia virus-T7 RNA polymerase expression system that when cells expressing the nucleocapsid protein (N), the phosphoprotein (NS), and the large polymerase protein (L) of VSV were superinfected with defective interfering (DI) particles, rapid and efficient replication and amplification of DI particle RNA occurred. Here, we demonstrate that all five VSV proteins can be expressed simultaneously when cells are cotransfected with plasmids containing the matrix protein (M) gene and the glycoprotein (G) gene of VSV in addition to plasmids containing the genes for the N, NS, and L proteins. When cells coexpressing all five VSV proteins were superinfected with DI particles, which because of their defectiveness are unable to express any viral proteins or to replicate, DI particle replication, assembly, and budding were observed and infectious DI particles were released into the culture fluids. Omission of either the M or G protein expression resulted in no DI particle budding. The vector-supported DI particles were similar in size and morphology to the authentic DI particles generated from cells coinfected with DI particles and helper VSV and their infectivity could be blocked by anti-VSV or anti-G antiserum. The successful replication, assembly, and budding of DI particles from cells expressing all five VSV proteins from cloned cDNAs provide a powerful approach for detailed structure-function analysis of the VSV gene products in each step of the replicative cycle of the virus.

Original languageEnglish (US)
Pages (from-to)1379-1383
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume88
Issue number4
DOIs
StatePublished - 1991

Keywords

  • Coexpression
  • Helper-independent budding
  • Vaccinia virus-T7 RNA polymerase

ASJC Scopus subject areas

  • General

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