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PMID: 3041041 Published · ppublish English Journal Article Research Support, U.S. Gov't, Non-P.H.S.

Proteolytic processing of foot-and-mouth disease virus polyproteins expressed in a cell-free system from clone-derived transcripts.

Journal of virology ·Vol. 61 ·No. 10 ·1987-10-00 ·Pages 3199-207

Vakharia VN, Devaney MA, Moore DM, Dunn JJ, Grubman MJ

Abstract

All picornaviral genes are expressed as a single, large polyprotein, which is proteolytically processed into the system produces functional proteins, including viral protease 3C, which plays a major role in processing the precursor proteins. To study the function of the two putative proteases 3C and leader (L) in processing, we constructed several cDNA plasmids encoding various regions of the FMDV type A12 genome. These plasmids, containing FMDV cDNA segments under the control of the T7 promoter, were transcribed in vitro by using T7 RNA polymerase and then translated in rabbit reticulocyte lysates. The expressed FMDV gene products were identified by immunoprecipitation with specific antisera and analyzed by gel electrophoresis. The results demonstrate the following: (i) the leader protein, L, is processed from the structural protein precursor, P1, in the absence of any P2 or P3 region proteins; (ii) protein 2A remains associated with the structural protein precursor, P1, rather than the precursor, P2; (iii) the processing of the P1-2A/P2 junction is not catalyzed by 3C or L; (iv) the proteolytic processing of polyproteins from the structural P1 region (except VP4/VP2) and the nonstructural P2 and P3 region is catalyzed by 3C.

MeSH Terms
Animals Aphthovirus/genetics,metabolism Cloning, Molecular DNA Restriction Enzymes DNA, Viral/genetics Deoxyribonucleases, Type II Site-Specific Genes, Viral Peptide Hydrolases/genetics,metabolism Plasmids Protein Biosynthesis Protein Processing, Post-Translational RNA, Viral/genetics Transcription, Genetic Viral Proteins/genetics,metabolism
Chemicals
DNA, Viral RNA, Viral Viral Proteins DNA Restriction Enzymes Deoxyribonucleases, Type II Site-Specific GATATC-specific type II deoxyribonucleases Peptide Hydrolases
Authors & Affiliations
5 authors, click to expand affiliations / ORCID
Vakharia V N
Devaney M A
Moore D M
Dunn J J
Grubman M J
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48 references, click to expand
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Article Info
Journal
Journal of virology
Abbr.
J Virol
ISSN
0022-538X
DOI
10.1128/JVI.61.10.3199-3207.1987
Published
1987-10-00
Pages
3199-207
Language
English
Region
United States
NLM ID
0113724
PMCID
PMC255898
Subset
IM
External Links
PubMed source DOI: 10.1128/JVI.61.10.3199-3207.1987 PMC Full Text View journal details
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