skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development

Abstract

ABSTRACT The foot-and-mouth disease virus (FMDV) afflicts livestock in more than 80 countries, limiting food production and global trade. Production of foot-and-mouth disease (FMD) vaccines requires cytosolic expression of the FMDV 3C protease to cleave the P1 polyprotein into mature capsid proteins, but the FMDV 3C protease is toxic to host cells. To identify less-toxic isoforms of the FMDV 3C protease, we screened 3C mutants for increased transgene output in comparison to wild-type 3C using aGaussialuciferase reporter system. The novel point mutation 3C(L127P) increased yields of recombinant FMDV subunit proteins in mammalian and bacterial cells expressing P1-3C transgenes and retained the ability to process P1 polyproteins from multiple FMDV serotypes. The 3C(L127P) mutant produced crystalline arrays of FMDV-like particles in mammalian and bacterial cells, potentially providing a practical method of rapid, inexpensive FMD vaccine production in bacteria. IMPORTANCEThe mutant FMDV 3C protease L127P significantly increased yields of recombinant FMDV subunit antigens and produced virus-like particles in mammalian and bacterial cells. The L127P mutation represents a novel advancement for economical FMD vaccine production.

Authors:
 [1];  [2];  [2];  [3];  [4];  [3];  [3];  [5];  [4];  [6];  [5];  [5]
  1. US Dept. of Homeland Security (DHS), Greenport, NY (United States). Plum Island Animal Disease Center; Leidos, Inc., Greenport, NY (United States). Plum Island Animal Disease Center; Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States). Plum Island Animal Disease Center Research Participation Program
  2. Leidos, Inc., Greenport, NY (United States). Plum Island Animal Disease Center; Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States). Plum Island Animal Disease Center Research Participation Program
  3. Leidos, Inc., Greenport, NY (United States). Plum Island Animal Disease Center
  4. Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States). Plum Island Animal Disease Center Research Participation Program
  5. US Dept. of Homeland Security (DHS), Greenport, NY (United States). Plum Island Animal Disease Center;
  6. Leidos, Inc., Greenport, NY (United States). Plum Island Animal Disease Center;
Publication Date:
Research Org.:
Oak Ridge Associated Univ., Oak Ridge, TN (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1626072
Grant/Contract Number:  
AC05-06OR23100; HSHQDC-14-F-00035; HSHQPM-14-X00001
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Journal of Virology
Additional Journal Information:
Journal Volume: 91; Journal Issue: 22; Journal ID: ISSN 0022-538X
Publisher:
American Society for Microbiology
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; 60 APPLIED LIFE SCIENCES; Virology; 3C protease; StopGo translation; Escherichia coli; Gaussia luciferase; VLP; adenoviruses; foot-and-mouth disease virus; in vivo; in vivo expression technology; translational interrupter; vaccines; virus-like particles

Citation Formats

Puckette, Michael, Clark, Benjamin A., Smith, Justin D., Turecek, Traci, Martel, Erica, Gabbert, Lindsay, Pisano, Melia, Hurtle, William, Pacheco, Juan M., Barrera, José, Neilan, John G., and Rasmussen, Max. Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development. United States: N. p., 2017. Web. doi:10.1128/jvi.00924-17.
Puckette, Michael, Clark, Benjamin A., Smith, Justin D., Turecek, Traci, Martel, Erica, Gabbert, Lindsay, Pisano, Melia, Hurtle, William, Pacheco, Juan M., Barrera, José, Neilan, John G., & Rasmussen, Max. Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development. United States. https://doi.org/10.1128/jvi.00924-17
Puckette, Michael, Clark, Benjamin A., Smith, Justin D., Turecek, Traci, Martel, Erica, Gabbert, Lindsay, Pisano, Melia, Hurtle, William, Pacheco, Juan M., Barrera, José, Neilan, John G., and Rasmussen, Max. Wed . "Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development". United States. https://doi.org/10.1128/jvi.00924-17. https://www.osti.gov/servlets/purl/1626072.
@article{osti_1626072,
title = {Foot-and-Mouth Disease (FMD) Virus 3C Protease Mutant L127P: Implications for FMD Vaccine Development},
author = {Puckette, Michael and Clark, Benjamin A. and Smith, Justin D. and Turecek, Traci and Martel, Erica and Gabbert, Lindsay and Pisano, Melia and Hurtle, William and Pacheco, Juan M. and Barrera, José and Neilan, John G. and Rasmussen, Max},
abstractNote = {ABSTRACT The foot-and-mouth disease virus (FMDV) afflicts livestock in more than 80 countries, limiting food production and global trade. Production of foot-and-mouth disease (FMD) vaccines requires cytosolic expression of the FMDV 3C protease to cleave the P1 polyprotein into mature capsid proteins, but the FMDV 3C protease is toxic to host cells. To identify less-toxic isoforms of the FMDV 3C protease, we screened 3C mutants for increased transgene output in comparison to wild-type 3C using aGaussialuciferase reporter system. The novel point mutation 3C(L127P) increased yields of recombinant FMDV subunit proteins in mammalian and bacterial cells expressing P1-3C transgenes and retained the ability to process P1 polyproteins from multiple FMDV serotypes. The 3C(L127P) mutant produced crystalline arrays of FMDV-like particles in mammalian and bacterial cells, potentially providing a practical method of rapid, inexpensive FMD vaccine production in bacteria. IMPORTANCEThe mutant FMDV 3C protease L127P significantly increased yields of recombinant FMDV subunit antigens and produced virus-like particles in mammalian and bacterial cells. The L127P mutation represents a novel advancement for economical FMD vaccine production.},
doi = {10.1128/jvi.00924-17},
url = {https://www.osti.gov/biblio/1626072}, journal = {Journal of Virology},
issn = {0022-538X},
number = 22,
volume = 91,
place = {United States},
year = {2017},
month = {9}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Citation Metrics:
Cited by: 12 works
Citation information provided by
Web of Science

Save / Share:

Works referenced in this record:

Foot-and-Mouth Disease Virus 3C Protease Induces Cleavage of Translation Initiation Factors eIF4A and eIF4G within Infected Cells
journal, January 2000


Crystallization of foot-and-mouth disease virus 3C protease: surface mutagenesis and a novel crystal-optimization strategy
journal, April 2005


Genetic heterogeneity in the foot-and-mouth disease virus Leader and 3C proteinases
journal, May 2002


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


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


Assembly and characterization of foot-and-mouth disease virus empty capsid particles expressed within mammalian cells
journal, August 2013


Electron microscopic observation of crystalline arrays of foot-and-mouth disease virus.
journal, January 1966


Biologically active protease of foot and mouth disease virus is expressed from cloned viral cDNA in Escherichia coli.
journal, June 1984


pAd5-Blue: Direct Ligation System for Engineering Recombinant Adenovirus Constructs
journal, November 2001


FMD vaccines
journal, January 2003


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


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


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


Foot-and-mouth disease virus protease 3C induces specific proteolytic cleavage of host cell histone H3.
journal, January 1990


Foot-and-Mouth Disease Virus 3C Protease Induces Fragmentation of the Golgi Compartment and Blocks Intra-Golgi Transport
journal, August 2013


Structural and Mutagenic Analysis of Foot-and-Mouth Disease Virus 3C Protease Reveals the Role of the β-Ribbon in Proteolysis
journal, October 2006


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


Characterization of the Foot-and-Mouth Disease Virus 3C Protease Expressed in Escherichia coli
journal, November 1993


Evaluation of a Fiber-Modified Adenovirus Vector Vaccine against Foot-and-Mouth Disease in Cattle
journal, November 2015


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


    Works referencing / citing this record: