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Virus derived from an Enterovirus B construct efficiently reverts from a frameshift mutation immediately beyond the translation initiation site
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences. (Picorna)ORCID iD: 0000-0003-4533-9094
Linnaeus University, Faculty of Health and Life Sciences, Department of Chemistry and Biomedical Sciences.
Linnaeus University, Faculty of Health and Life Sciences, Department of Biology and Environmental Science.ORCID iD: 0000-0002-1426-0036
The Pennsylvania State University College of Medicine, USA.
Show others and affiliations
(English)Manuscript (preprint) (Other academic)
National Category
Biochemistry and Molecular Biology
Research subject
Biomedical Sciences, Virology
Identifiers
URN: urn:nbn:se:lnu:diva-41540OAI: oai:DiVA.org:lnu-41540DiVA: diva2:800024
Available from: 2015-04-01 Created: 2015-04-01 Last updated: 2016-10-05Bibliographically approved
In thesis
1. Reverse genetic studies of Enterovirus replication
Open this publication in new window or tab >>Reverse genetic studies of Enterovirus replication
2015 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Enteroviruses belong to the Picornaviridae family and are small icosahedral viruses with RNA genomes of positive polarity, containing a single open reading frame. They mostly cause mild or asymptomatic infections, but also a wide array of diseases including: poliomyelitis, encephalitis, gastroenteritis, aseptic meningitis, myocarditis, hand-foot-and-mouth disease, hepatitis and respiratory diseases, ranging from severe infections to the common cold. The projects described in this thesis have been carried out through reverse genetic studies of Enterovirus B and Rhinovirus C.

                  In Papers I and II, a cassette vector was used to study recombination and translation of the RNA genome. It was found that the non-structural coding region could replicate when combined with the structural protein-coding region of other viruses of the same species. Furthermore, the genome could be translated and replicated without the presence of the structural protein-coding region. Moreover, it was found that when two additional nucleotides were introduced, shifting the reading frame, the virus could revert to the original reading frame, restoring efficient replication. In Paper III, a vector containing the genome of echovirus 5 was altered to produce an authentic 5’end of the in vitro transcribed RNA, which increased efficiency of replication initiation 20 times. This result is important, as it may lead to more efficient oncolytic virotherapy. An authentic 5’end was further used in Paper IV, where replication of Rhinovirus C in cell lines was attempted. Although passaging of the virus was unsuccessful, the genome was replicated and cytopathic effect induced after transfection. The restriction of efficient replication was therefore hypothesized to lie in the attachment and entry stages of the replication cycle. In Paper V, a cytolytic virus was found to have almost 10 times larger impact on gene expression of the host cell than a non-cytolytic variant. Furthermore, the lytic virus was found to build up inside the host cell, while the non-cytolytic virus was efficiently released.

                  As a whole, this thesis has contributed to a deeper understanding of replication of enteroviruses, which may prove important in development of novel vaccines, antiviral agents and oncolytic virotherapies.

Place, publisher, year, edition, pages
Växjö: Linnaeus University Press, 2015
Series
Linnaeus University Dissertations, 216/2015
Keyword
Enterovirus, picornavirus, coxsackievirus, echovirus, rhinovirus, reverse genetics, replication, translation, transfection, cassette vector.
National Category
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
Research subject
Biomedical Sciences, Virology
Identifiers
urn:nbn:se:lnu:diva-41636 (URN)978-91-87925-55-9 (ISBN)
Public defence
2015-04-24, N2007K, Smålandsgatan 26, Kalmar, 09:00 (English)
Opponent
Supervisors
Available from: 2015-04-10 Created: 2015-04-01 Last updated: 2015-04-10Bibliographically approved

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CiteExportLink to record
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