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Title: The biological role of viral tRNA-like molecules in a murine gammaherpesvirus infection
Author: Cliffe, A.
Awarding Body: University of Edinburgh
Current Institution: University of Edinburgh
Date of Award: 2006
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The aim of this project was to characterise the vtRNAs. The presence of the vtRNAs within purified, RNase treated viral stocks indicated their packaging within the MHV-68 virion. Although both viral and cellular mRNAs were also present, it appeared that the major RNA species packaged by MHV-68 were small RNA molecules, such as the vtRNAs. Incorporation of RNA molecules into the virion is not unique to MHV-68 as other herpesviruses have been found to package RNA, although the vtRNAs represent the only packaged small viral non-coding RNA molecules discovered so far. In addition, this is the first study to demonstrate the preferential incorporation of small RNA molecules by a herpesvirus. The mechanism by which the vtRNAs assemble into the virion is not clear. In situ hybridisation demonstrated that within infected cells the vtRNAs localised to globular areas within the nucleus and were also found at high levels within the cytoplasm. electrophoretic mobility shift assays performed using vtRNA1 and vtRNA4 indicated binding to protein complexes present within both the nucleus and cytoplasm of infected cells. Inhibition of vtRNA-protein binding by an anti-MHV-68 antibody indicated direct interaction of the vtRNAs with viral protein(s). Hence it is likely that their incorporation is mediated through binding to viral protein(s) during virion assembly in either the nucleus or cytoplasm. MHV-76 is a deletion mutant of MHV-68, which lacks all eight vtRNAs along with four other genes (M1-M4). The contribution of the vtRNAs to viral pathogenesis has been investigated by construction of recombinant MHV-76 which expressed vtRNAs1-5 under their natural promoters. The recombinant viruses produced in this project will provide excellent tools to investigate their function further through both in vitro and in vivo analysis.
Supervisor: Not available Sponsor: Not available
Qualification Name: Thesis (Ph.D.) Qualification Level: Doctoral
EThOS ID:  DOI: Not available