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PMID: 8627754 Published · ppublish English Journal Article

Vaccinia virus A17L open reading frame encodes an essential component of nascent viral membranes that is required to initiate morphogenesis.

Journal of virology ·Vol. 70 ·No. 5 ·1996-05-00 ·Pages 2797-808

Wolffe EJ, Moore DM, Peters PJ, Moss B

Abstract

We generated an antiserum to the predicted C-terminal peptide of the A17L open reading frame (ORF), which encodes a 23-kDa polypeptide with hydrophobic regions characteristic of membrane proteins. Immuno-electron microscopy of infected cells indicated that the A17L protein is intimately associated with the earliest characteristic viral membranes, even those formed in the presence of the drug rifampin. To study the role of the A17L protein in morphogenesis, we constructed recombinant vaccinia viruses in which the endogenous A17L ORF was deleted and a copy of the ORF under the control of the bacteriophage T7 RNA polymerase and the Escherichia coli lac repressor was inserted into an alternative site in the vaccinia virus genome. Growth of these recombinant viruses was entirely dependent on the induction of A17L expression by isopropyl-beta-D-thiogalactopyranoside. Electron microscopic examination of cells infected in the absence of inducer revealed the accumulation of large, well-demarcated electron-dense aggregates but no characteristic membrane-associated viral structures. Viral late protein synthesis occurred under these conditions, although the maturational proteolytic processing of structural proteins was inhibited. We conclude that the product of the A17L gene is an essential component of the immature viral membrane and has an early function in viral morphogenesis.

MeSH Terms
Animals Base Sequence Blotting, Southern Cell Line Chlorocebus aethiops DNA Primers DNA, Viral/metabolism Genetic Vectors HeLa Cells Herpesvirus 3, Human/genetics,physiology,ultrastructure Humans Isopropyl Thiogalactoside/pharmacology Membrane Proteins Microscopy, Electron Microscopy, Immunoelectron Molecular Sequence Data Morphogenesis/drug effects Open Reading Frames Plasmids Polymerase Chain Reaction Restriction Mapping Viral Envelope Proteins/analysis,genetics,metabolism Viral Plaque Assay Virus Replication
Chemicals
A17L protein, Vaccinia virus DNA Primers DNA, Viral Membrane Proteins Viral Envelope Proteins Isopropyl Thiogalactoside
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Wolffe E J
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, Bethesda, Maryland 20892, USA.
Moore D M
Peters P J
Moss B
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.70.5.2797-2808.1996
Published
1996-05-00
Pages
2797-808
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC190137
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.70.5.2797-2808.1996 PMC Full Text View journal details
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