Next Article in Journal
Modelling Degradation and Replication Kinetics of the Zika Virus In Vitro Infection
Previous Article in Journal
Antibiofilm Activity of a Broad-Range Recombinant Endolysin LysECD7: In Vitro and In Vivo Study
Previous Article in Special Issue
The Role of Temperature in Transmission of Zoonotic Arboviruses
Open AccessArticle

Measuring Alphavirus Fidelity Using Non-Infectious Virus Particles

1
Department of Pathology, University of Texas Medical Branch, Galveston, TX 77555, USA
2
Centre for Neglected Tropical Diseases, Departments of Vector Biology and Tropical Disease Biology, Liverpool School of Tropical Medicine, Pembroke Place, Liverpool L3 5QA, UK
3
Department of Pharmacology and Toxicology, Sealy Center for Structural Biology and Molecular Biophysics, University of Texas Medical Branch, Galveston, TX 77555, USA
4
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine, University of California, Davis, Davis, CA 95616, USA
5
Department of Microbiology and Immunology, University of Texas Medical Branch, Galveston, TX 77555, USA
*
Author to whom correspondence should be addressed.
Current address: Glenn Biggs Institute for Alzheimer’s and Neurodegenerative Diseases, UT Health San Antonio, San Antonio, TX 78229, USA.
Current address: Department of Microbiology and Immunology, SUNY Center for Environmental Health and Medicine, Institute for Global Health and Translational Science, SUNY Upstate Medical University, Syracuse, NY 13210, USA.
§
Current address: School of Life Sciences, University of Keele, Keele ST5 5BG, UK.
Viruses 2020, 12(5), 546; https://doi.org/10.3390/v12050546
Received: 3 April 2020 / Revised: 6 May 2020 / Accepted: 13 May 2020 / Published: 15 May 2020
(This article belongs to the Special Issue Transmission Dynamics of Insect Viruses)
Mutations are incorporated into the genomes of RNA viruses at an optimal frequency and altering this precise frequency has been proposed as a strategy to create live-attenuated vaccines. However, determining the effect of specific mutations that alter fidelity has been difficult because of the rapid selection of the virus population during replication. By deleting residues of the structural polyprotein PE2 cleavage site, E3Δ56-59, in Venezuelan equine encephalitis virus (VEEV) TC-83 vaccine strain, non-infectious virus particles were used to assess the effect of single mutations on mutation frequency without the interference of selection that results from multiple replication cycles. Next-generation sequencing analysis revealed a significantly lower frequency of transversion mutations and overall mutation frequency for the fidelity mutants compared to VEEV TC-83 E3Δ56-59. We demonstrate that deletion of the PE2 cleavage site halts virus infection while making the virus particles available for downstream sequencing. The conservation of the site will allow the evaluation of suspected fidelity mutants across alphaviruses of medical importance. View Full-Text
Keywords: alphavirus; arbovirus; fidelity mutants; mutation frequency alphavirus; arbovirus; fidelity mutants; mutation frequency
Show Figures

Figure 1

MDPI and ACS Style

Patterson, E.I.; Khanipov, K.; Swetnam, D.M.; Walsdorf, S.; Kautz, T.F.; Thangamani, S.; Fofanov, Y.; Forrester, N.L. Measuring Alphavirus Fidelity Using Non-Infectious Virus Particles. Viruses 2020, 12, 546.

Show more citation formats Show less citations formats
Note that from the first issue of 2016, MDPI journals use article numbers instead of page numbers. See further details here.

Article Access Map by Country/Region

1
Search more from Scilit
 
Search
Back to TopTop