Absence of E protein arrests transmissible gastroenteritis coronavirus maturation in the secretory pathway
- Centro Nacional de Biotecnologia, CSIC, Department of Molecular and Cell Biology, Campus Universidad Autonoma, Darwin 3, Cantoblanco, 28049 Madrid (Spain)
- Fort-Dodge Veterinaria, Department of Research and Development, Girona (Spain)
- Centro Nacional de Biotecnologia, CSIC, Macromolecular Structure, Campus Universidad Autonoma, Darwin 3, Cantoblanco, 28049 Madrid (Spain)
A recombinant transmissible gastroenteritis coronavirus (rTGEV) in which E gene was deleted (rTGEV-{delta}E) has been engineered. This deletion mutant only grows in cells expressing E protein (E{sup +} cells) indicating that E was an essential gene for TGEV replication. Electron microscopy studies of rTGEV-{delta}E infected BHK-pAPN-E{sup -} cells showed that only immature intracellular virions were assembled. These virions were non-infectious and not secreted to the extracellular medium in BHK-pAPN-E{sup -} cells. RNA and protein composition analysis by RNase-gold and immunoelectron microscopy showed that rTGEV-{delta}E virions contained RNA and also all the structural TGEV proteins, except the deleted E protein. Nevertheless, full virion maturation was blocked. Studies of the rTGEV-{delta}E subcellular localization by confocal and immunoelectron microscopy in infected E{sup -} cells showed that in the absence of E protein virus trafficking was arrested in the intermediate compartment. Therefore, the absence of E protein in TGEV resulted in two actions, a blockade of virus trafficking in the membranes of the secretory pathway, and prevention of full virus maturation.
- OSTI ID:
- 21077994
- Journal Information:
- Virology, Journal Name: Virology Journal Issue: 2 Vol. 368; ISSN VIRLAX; ISSN 0042-6822
- Country of Publication:
- United States
- Language:
- English
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