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

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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.

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Authors and Affiliations

  1. Institute of Molecular and Cellular Virology, Federal Research Centre for Virus Diseases of Animals, Friedrich-Loeffler-Institutes, D-17498, Insel Riems, Germany

    Siegfried Weber

  2. Institute of Diagnostic Virology, Federal Research Centre for Virus Diseases of Animals, Friedrich-Loeffler-Institutes, D-17498, Insel Riems, Germany

    Harald Granzow

  3. Institute of Microbiology, D-72076, Tübingen, Germany

    Frank Weiland & Otfried Marquardt

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  1. Siegfried Weber
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  2. Harald Granzow
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Weber, S., Granzow, H., Weiland, F. et al. Intracellular membrane proliferation in E. coli induced by foot-and-mouth disease virus 3A gene products. Virus Genes 12, 5–14 (1996). https://doi.org/10.1007/BF00369995

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  • DOI: https://doi.org/10.1007/BF00369995

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