Absence of E protein arrests transmissible gastroenteritis coronavirus maturation in the secretory pathway.
A recombinant transmissible gastroenteritis coronavirus (rTGEV) in which E gene was deleted (rTGEV-DeltaE) has been engineered. This deletion mutant only grows in cells expressing E protein (E(+) cells) indicating that E was an essential gene for TGEV replication. Electron microscopy studies of rTGEV-DeltaE infected BHK-pAPN-E(-) 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(-) cells. RNA and protein composition analysis by RNase-gold and immunoelectron microscopy showed that rTGEV-DeltaE 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-DeltaE subcellular localization by confocal and immunoelectron microscopy in infected E(-) 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.
Ortego
Javier
J
Centro Nacional de Biotecnología, CSIC, Department of Molecular and Cell Biology, Campus Universidad Autónoma, Darwin 3, Cantoblanco, 28049 Madrid, Spain.
Ceriani
Juan E
JE
Patiño
Cristina
C
Plana
Juan
J
Enjuanes
Luis
L
D000818:Animals; D002460:Cell Line; D006224:Cricetinae; D017353:Gene Deletion; D020043:Genes, Essential; D018374:LLC-PK1 Cells / Q000821:virology; D013552:Swine; D005760:Transmissible gastroenteritis virus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism / Q000502:physiology*; D014759:Viral Envelope Proteins / Q000235:genetics* / Q000378:metabolism; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism; D014771:Virion / Q000254:growth & development / Q000378:metabolism; D014779:Virus Replication
D014759:Viral Envelope Proteins; D014764:Viral Proteins
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