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Virus Genes

, Volume 12, Issue 1, pp 5–14 | Cite as

Intracellular membrane proliferation in E. coli induced by foot-and-mouth disease virus 3A gene products

  • Siegfried Weber
  • Harald Granzow
  • Frank Weiland
  • Otfried Marquardt
Article

Abstract

During picornavirus infection replication of genomic RNA occurs in membrane-associated ribonucleoprotein complexes. These replication complexes contain different nonstructural viral proteins with mostly unknown function. To examine the function of nonstructural picornaviral proteins in more detail, cDNA of foot-and-mouth-disease virus (FMDV) strain O1 Lausanne was cloned into lambda ZAP II, and different parts of the P3-coding sequence were expressed in E. coli by the T7 polymerase system. Expression products constituted (a) fusion proteins composed of N-terminal leader peptide of bacteriophage T7 π10 protein fused to FMDV P3-sequences of different lengths, (b) translation products of authentic P3-region genes, and (c) carboxy-terminally truncated 3A proteins. Expression products were characterized by NaDodSO4-polyacrylamide gel electrophoresis, immunoblotting, as well as electron and immunoelectron microscopy. We show here that in the T7 polymerase system a high level of expression of 3A-containing peptides is achieved in E. coli. Remarkably, the expression of 3A-derived proteins induced a dramatic intracellular membrane proliferation in E. coli cells, similar to the vesicle induction observed in FMDV-infected cells. By immunoelectron microscopy, 3A-reactive material was found associated with these membranes. We hypothesize that the FMDV 3A protein is instrumental in eliciting intracellular membrane proliferation in infected cells as a prerequisite for viral RNA replication.

Key words

picornavirus foot-and-mouth disease virus FMDV T7 polymerase 3A gene product membrane proliferation 

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Copyright information

© Kluwer Academic Publishers 1996

Authors and Affiliations

  • Siegfried Weber
    • 1
  • Harald Granzow
    • 2
  • Frank Weiland
    • 3
  • Otfried Marquardt
    • 3
  1. 1.Institute of Molecular and Cellular VirologyFederal Research Centre for Virus Diseases of Animals, Friedrich-Loeffler-InstitutesInsel RiemsGermany
  2. 2.Institute of Diagnostic VirologyFederal Research Centre for Virus Diseases of Animals, Friedrich-Loeffler-InstitutesInsel RiemsGermany
  3. 3.Institute of MicrobiologyTübingenGermany

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