Journal article
The hepatitis A virus polyprotein expressed by a recombinant vaccinia virus undergoes proteolytic processing and assembly into viruslike particles
Journal of virology, Vol.65(9), pp.5029-5036
09/1991
DOI: 10.1128/JVI.65.9.5029-5036.1991
PMCID: PMC248966
PMID: 1651421
Abstract
Hepatitis A virus (HAV) contains a single-stranded, plus-sense RNA genome with a single long open reading frame encoding a polyprotein of approximately 250 kDa. Viral structural proteins are generated by posttranslational proteolytic processing of this polyprotein. We constructed recombinant vaccinia viruses which expressed the HAV polyprotein (rV-ORF) and the P1 structural region (rV-P1). rV-ORF-infected cell lysates demonstrated that the polyprotein was cleaved into immunoreactive 29- and 33-kDa proteins which comigrated with HAV capsid proteins VP0 and VP1. The rV-P1 construct produced a 90-kDa protein which showed no evidence of posttranslational processing. Solid-phase radioimmunoassays with human polyclonal anti-HAV sera and with murine or human neutralizing monoclonal anti-HAV antibodies recognized the rV-ORF-infected cell lysates. Sucrose density gradients of rV-ORF-infected cell lysates contained peaks of HAV antigen with sedimentation coefficients of approximately 70S and 15S, similar to those of HAV empty capsids and pentamers. Immune electron microscopy also demonstrated the presence of viruslike particles in rV-ORF-infected cell lysates. Thus, the HAV polyprotein expressed by a recombinant vaccinia virus demonstrated posttranslational processing into mature capsid proteins which assembled into antigenic viruslike particles.
Details
- Title: Subtitle
- The hepatitis A virus polyprotein expressed by a recombinant vaccinia virus undergoes proteolytic processing and assembly into viruslike particles
- Creators
- P L Winokur - Department of Internal Medicine, University of Iowa College of Medicine, Iowa CityJ H McLinden - Department of Internal Medicine, University of Iowa College of Medicine, Iowa CityJ T Stapleton - Department of Internal Medicine, University of Iowa College of Medicine, Iowa City
- Resource Type
- Journal article
- Publication Details
- Journal of virology, Vol.65(9), pp.5029-5036
- DOI
- 10.1128/JVI.65.9.5029-5036.1991
- PMID
- 1651421
- PMCID
- PMC248966
- ISSN
- 0022-538X
- eISSN
- 1098-5514
- Language
- English
- Date published
- 09/1991
- Academic Unit
- Microbiology and Immunology; Infectious Diseases; Medicine Administration; Internal Medicine
- Record Identifier
- 9984094317502771
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