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Cells that express all five proteins of vesicular stomatitis virus from cloned cDNAs support replication, assembly,

A K Pattnaik1, G W Wertz

  • 1Department of Microbiology, University of Alabama, Birmingham Medical School 35294.

Proceedings of the National Academy of Sciences of the United States of America
|February 15, 1991
PubMed

Insights

Researchers developed a new method to study vesicular stomatitis virus (VSV) proteins. This approach allows for the analysis of all five VSV proteins, enabling a deeper understanding of viral replication and gene function.

Area of Science:

  • Virology
  • Molecular Biology
  • Biochemistry

Background:

  • Vesicular stomatitis virus (VSV) structure-function analysis is crucial for understanding viral replication.
  • Previous studies utilized the vaccinia virus-T7 RNA polymerase system to express VSV N, NS, and L proteins, facilitating defective interfering (DI) particle RNA replication.

Purpose of the Study:

  • To establish an alternative approach for VSV structure-function analysis.
  • To investigate the role of all five VSV proteins in the viral life cycle.
  • To demonstrate the successful replication, assembly, and budding of DI particles using coexpressed VSV proteins.

Main Methods:

  • Co-transfection of cells with plasmids encoding all five VSV proteins (N, NS, L, M, G).
  • Superinfection of expressing cells with VSV defective interfering (DI) particles.
  • Analysis of DI particle replication, assembly, and budding.
  • Characterization of vector-supported DI particles for size, morphology, and infectivity.

Main Results:

  • Simultaneous expression of all five VSV proteins was achieved.
  • Coexpression enabled DI particle replication, assembly, and budding, releasing infectious DI particles.
  • Omission of matrix (M) or glycoprotein (G) protein expression abolished DI particle budding.
  • Vector-supported DI particles mimicked authentic DI particles in characteristics and infectivity.

Conclusions:

  • The coexpression system provides a powerful platform for VSV structure-function analysis.
  • This method allows detailed investigation of VSV gene products at each stage of the viral replicative cycle.
  • The M and G proteins are essential for DI particle budding.

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