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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

Journal Article · · Journal of General Virology
DOI:https://doi.org/10.1099/jgv.0.001204· OSTI ID:1580726
 [1];  [2];  [3];  [4];  [5];  [5]
  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)
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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.

Research Organization:
Oak Ridge Inst. for Science and Education (ORISE), Oak Ridge, TN (United States)
Sponsoring Organization:
USDOE
Grant/Contract Number:
SC0014664; AC05-06OR23100
OSTI ID:
1580726
Journal Information:
Journal of General Virology, Vol. 100, Issue 3; ISSN 0022-1317
Publisher:
Microbiology SocietyCopyright Statement
Country of Publication:
United States
Language:
English
Citation Metrics:
Cited by: 3 works
Citation information provided by
Web of Science

References (23)

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

59 BASIC BIOLOGICAL SCIENCES
FMDV
proteolysis
Vaccine
structure
mutagenesis
3C