Identification and characterization of cell-adapted mutations in West Nile Virus and replicons

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Identification and characterization of cell-adapted mutations in West Nile Virus and replicons

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dc.contributor.advisor Peter W. Mason en_US
dc.creator Shannan Lee Rossi en_US
dc.date.accessioned 2011-12-20T16:04:13Z
dc.date.available 2008-04-03 en_US
dc.date.available 2011-12-20T16:04:13Z
dc.date.created 2008-02-27 en_US
dc.date.issued 2007-12-19 en_US
dc.identifier.other etd-02272008-070728 en_US
dc.identifier.uri http://hdl.handle.net/2152.3/32
dc.description.abstract Flavivirus persistence in cell culture is achieved by altering the interaction between the viral genome and the host cell. To identify some of the factors that contribute to a persistent West Nile virus (WNV) infection, WNV subgenomic replicon (WNR) genomes were modified to contain neomycin phosphotransferase (antibiotic-resistance) gene and cells that persistently harbored this WNR were selected for using G418. There were many changes to the genomes harvested from WNR-bearing cell cultures compared to the parental genome, perhaps the most striking was the prevalence of NS2A mutations. WNR and WNV genomes harboring these mutations replicated more poorly than wt WNR or WNV genomes in vitro. These NS2A mutant genomes, as well as a genome with a large deletion in the 3’ UTR, were highly attenuated Swiss-Webster outbred mice. Low levels of IFN were produced from cells infected with NS2A or 3’ UTR deletion WNR genomes compared to wt WNR genomes. Since the WNR and WNV cannot efficiently replicate, little stress is placed on the cell during replication, resulting in minimal engagement of the cell’s stress and apoptotic responses and a noncytopathic infection. As a result, little cytopathic effect and apoptotic death were observed in cells infected with NS2A or 3’ UTR deletion WNR mutants compared to cells infected with wt WNR genomes. This lack of death may have been attributed to low levels of CHOP, a pro-apoptotic protein that is induced during endoplasmic reticulum stress. Taken together, these data support the hypothesis that WNR and WNV genomes that replicate poorly do not efficiently produce signs of their replication that can be recognized by the cell and place little stress upon the cell, resulting in an attenuated noncytopathic, persistent infection. en_US
dc.format.medium electronic en_US
dc.language.iso eng en_US
dc.rights Copyright © is held by the author. Presentation of this material on the TDL web site by The University of Texas Medical Branch at Galveston was made possible under a limited license grant from the author who has retained all copyrights in the works. en_US
dc.subject West Nile virus en_US
dc.subject persistence en_US
dc.subject interferon en_US
dc.subject Flavivirus en_US
dc.subject ER stress en_US
dc.subject attenuation en_US
dc.title Identification and characterization of cell-adapted mutations in West Nile Virus and replicons en_US
dc.type.material text en_US
dc.type.genre dissertation en_US
thesis.degree.name PhD en_US
thesis.degree.level Doctoral en_US
thesis.degree.grantor The University of Texas Medical Branch en_US
thesis.degree.department Experimental Pathology en_US
dc.contributor.committeeMember Scott Weaver en_US
dc.contributor.committeeMember Robert Tesh en_US
dc.contributor.committeeMember Ilya Frolov en_US
dc.contributor.committeeMember Charles Rice en_US

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