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Title: Effect of foot-and-mouth disease virus 3C protease B2 β-strand proline mutagenesis on expression and processing of the P1 polypeptide using a plasmid expression vector

Abstract

The production of experimental molecular vaccines against foot-and-mouth disease virus utilizes the viral encoded 3C protease for processing of the P1 polyprotein. Expression of wild type 3C protease is detrimental to host cells. The molecular vaccine constructs containing the 3C protease L127P mutant significantly reduce adverse effects associated with protease expression while retaining the ability to process and assemble virus-like particles. In published 3C protease crystal structures, the L127 residue is contained within the B2 β-strand as part of the A2–B2 β-sheet. To provide insight into the mechanism by which the L127P mutant alters the properties of the 3C protease, we performed scanning proline mutagenesis of residues 123–128 of the B2 β-strand and monitored expression and P1 processing. Simultaneously, we utilized random mutagenesis of the full 3C sequence to identify additional mutations presenting a phenotype similar to the L127P mutation. Six of the tested mutants enhanced expression over wild type, and the I22P, T100P and V124P mutations surpassed the L127P mutation in certain cell lines. These data are interpreted in conjunction with published 3C protease crystal structures to provide insight into the mechanism by which these mutations enhance expression.

Authors:
 [1];  [2];  [3];  [4];  [5];  [5]
+ Show Author Affiliations
  1. Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States)
  2. Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States); Coastal Carolina Univ., Conway, SC (United States)
  3. Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States); Bard High School Early College Baltimore, Baltimore, MD (United States)
  4. Leidos, Inc., Greenport, NY (United States)
  5. U.S. Department of Homeland Security Science and Technology Directorate, Greenport, NY (United States)
Publication Date:
Research Org.:
Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1580726
Grant/Contract Number:  
SC0014664; AC05-06OR23100
Resource Type:
Accepted Manuscript
Journal Name:
Journal of General Virology
Additional Journal Information:
Journal Volume: 100; Journal Issue: 3; Journal ID: ISSN 0022-1317
Publisher:
Microbiology Society
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; FMDV; proteolysis; Vaccine; structure; mutagenesis; 3C

Citation Formats

Martel, Erica, Forzono, Emily, Kurker, Richard, Clark, Benjamin A., Neilan, John G., and Puckette, Michael. Effect of foot-and-mouth disease virus 3C protease B2 β-strand proline mutagenesis on expression and processing of the P1 polypeptide using a plasmid expression vector. United States: N. p., 2019. Web. doi:10.1099/jgv.0.001204.
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Martel, Erica, Forzono, Emily, Kurker, Richard, Clark, Benjamin A., Neilan, John G., & Puckette, Michael. Effect of foot-and-mouth disease virus 3C protease B2 β-strand proline mutagenesis on expression and processing of the P1 polypeptide using a plasmid expression vector. United States. https://doi.org/10.1099/jgv.0.001204
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Martel, Erica, Forzono, Emily, Kurker, Richard, Clark, Benjamin A., Neilan, John G., and Puckette, Michael. Thu . "Effect of foot-and-mouth disease virus 3C protease B2 β-strand proline mutagenesis on expression and processing of the P1 polypeptide using a plasmid expression vector". United States. https://doi.org/10.1099/jgv.0.001204. https://www.osti.gov/servlets/purl/1580726.
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@article{osti_1580726,
title = {Effect of foot-and-mouth disease virus 3C protease B2 β-strand proline mutagenesis on expression and processing of the P1 polypeptide using a plasmid expression vector},
author = {Martel, Erica and Forzono, Emily and Kurker, Richard and Clark, Benjamin A. and Neilan, John G. and Puckette, Michael},
abstractNote = {The production of experimental molecular vaccines against foot-and-mouth disease virus utilizes the viral encoded 3C protease for processing of the P1 polyprotein. Expression of wild type 3C protease is detrimental to host cells. The molecular vaccine constructs containing the 3C protease L127P mutant significantly reduce adverse effects associated with protease expression while retaining the ability to process and assemble virus-like particles. In published 3C protease crystal structures, the L127 residue is contained within the B2 β-strand as part of the A2–B2 β-sheet. To provide insight into the mechanism by which the L127P mutant alters the properties of the 3C protease, we performed scanning proline mutagenesis of residues 123–128 of the B2 β-strand and monitored expression and P1 processing. Simultaneously, we utilized random mutagenesis of the full 3C sequence to identify additional mutations presenting a phenotype similar to the L127P mutation. Six of the tested mutants enhanced expression over wild type, and the I22P, T100P and V124P mutations surpassed the L127P mutation in certain cell lines. These data are interpreted in conjunction with published 3C protease crystal structures to provide insight into the mechanism by which these mutations enhance expression.},
doi = {10.1099/jgv.0.001204},
journal = {Journal of General Virology},
number = 3,
volume = 100,
place = {United States},
year = {Thu Jan 31 00:00:00 EST 2019},
month = {Thu Jan 31 00:00:00 EST 2019}
}
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Journal Article:
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Publisher's Version of Record
https://doi.org/10.1099/jgv.0.001204
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Works referenced in this record:

Immune responses and protection against foot-and-mouth disease virus (FMDV) challenge in swine vaccinated with adenovirus-FMDV constructs
journal, February 2001

Identification of the Active-Site Residues of the 3C Proteinase of Foot-and-Mouth Disease Virus
journal, November 1995

  • Grubman, Marvin J.; Zellner, Marla; Bablanian, Gayne
  • Virology, Vol. 213, Issue 2
  • DOI: 10.1006/viro.1995.0030

Insights into Cleavage Specificity from the Crystal Structure of Foot-and-Mouth Disease Virus 3C Protease Complexed with a Peptide Substrate
journal, January 2010

  • Zunszain, Patricia A.; Knox, Stephen R.; Sweeney, Trevor R.
  • Journal of Molecular Biology, Vol. 395, Issue 2
  • DOI: 10.1016/j.jmb.2009.10.048

Production of Foot-and-mouth Disease Virus Antigen from BHK 21 Clone 13 Cells grown and infected in Deep Suspension Cultures
journal, March 1965

  • Capstick, P. B.; Garland, A. J.; Chapman, W. G.
  • Nature, Vol. 205, Issue 4976
  • DOI: 10.1038/2051135a0

Crystal structure of the 3C protease from Southern African Territories type 2 foot-and-mouth disease virus
journal, January 2016

Foot-and-Mouth Disease Virus 3C Protease Cleaves NEMO To Impair Innate Immune Signaling
journal, June 2012

Protein Coexpression Using FMDV 2A: Effect of “Linker” Residues
journal, January 2013

  • Minskaia, Ekaterina; Ryan, Martin D.
  • BioMed Research International, Vol. 2013
  • DOI: 10.1155/2013/291730

The economic impacts of foot and mouth disease – What are they, how big are they and where do they occur?
journal, November 2013

Growth of Foot-and-Mouth Disease Virus in a Fibroblastic Cell Line Derived from Hamster Kidneys
journal, April 1962

  • Mowat, G. N.; Chapman, W. G.
  • Nature, Vol. 194, Issue 4825
  • DOI: 10.1038/194253a0

Crystal Structure of Foot-and-Mouth Disease Virus 3C Protease: NEW INSIGHTS INTO CATALYTIC MECHANISM AND CLEAVAGE SPECIFICITY
journal, January 2005

  • Birtley, J. R.; Knox, S. R.; Jaulent, A. M.
  • Journal of Biological Chemistry, Vol. 280, Issue 12
  • DOI: 10.1074/jbc.M413254200

Gaussia luciferase reporter assay for monitoring biological processes in culture and in vivo
journal, April 2009

The nuclear protein Sam68 is cleaved by the FMDV 3C protease redistributing Sam68 to the cytoplasm during FMDV infection of host cells
journal, March 2012

Analysis of Foot-and-Mouth Disease Virus Type O1 Brugge Neutralization Epitopes Using Monoclonal Antibodies
journal, October 1986

Production and characterization of two serotype independent monoclonal antibodies against foot-and-mouth disease virus
journal, January 2007

  • Yang, Ming; Clavijo, Alfonso; Suarez-Banmann, Rachel
  • Veterinary Immunology and Immunopathology, Vol. 115, Issue 1-2
  • DOI: 10.1016/j.vetimm.2006.10.002

Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development
journal, September 2017

  • Puckette, Michael; Clark, Benjamin A.; Smith, Justin D.
  • Journal of Virology, Vol. 91, Issue 22
  • DOI: 10.1128/JVI.00924-17

Efficient production of foot-and-mouth disease virus empty capsids in insect cells following down regulation of 3C protease activity
journal, February 2013

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