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The function of extensive structured RNA in the evasion of host anti-virus responses
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Hornsey, Crystal A. (2012) The function of extensive structured RNA in the evasion of host anti-virus responses. PhD thesis, University of Warwick.
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Official URL: http://webcat.warwick.ac.uk/record=b2681731~S1
Abstract
Genome scale ordered RNA structure (GORS) is found throughout the genome of
many single stranded positive sense RNA viruses, including plant viruses. It was
hypothesised that GORS may function to help evade RNAi either by preventing the
generation of siRNAs or by stopping RNAi-mediated cleavage of the target. This
project used Potato Leafroll Virus (PLRV) to investigate the function of GORS in
plant viruses.
The RNA structure of a 750nt region of the genome was modified to have less, more
or the same energy as the WT sequence. The physical structure of these sequences
was shown to be different using two distinct methods. Viral infectivity was tested
and although all four viruses were able to replicate and spread to distal leaves, the
WT virus was always able to outcompete the variant viruses in competition assays.
This suggests GORS provides a distinct selective advantage to the WT virus.
The effect on the siRNA response was tested using a dedicated siRNA assay. In
plants, this showed that the WT sequence was more resistant to degradation by
siRNAs than the variant sequences in the presence of their specific inducers. The WT
inducer was also not able to cause suppression of the other targets indicating that this
inducer failed to produce siRNAs or that they were not effective. The siRNA
populations generated during infections were sequenced and the profiles compared.
This showed that all four viruses stimulated the production of siRNAs but the
location of siRNA hotspots differed. It is therefore hypothesised that GORS may
function to evade the RNAi response by directing the generation of less effective
siRNAs.
The data presented in this thesis not only informs current work on GORS and RNA
structure in viral genomes but also has wider implications for research on siRNAs
and food biosecurity.
Item Type: | Thesis (PhD) | ||||
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Subjects: | Q Science > QP Physiology Q Science > QR Microbiology > QR355 Virology |
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Library of Congress Subject Headings (LCSH): | RNA viruses, RNA -- Structure, Viral genomes, Host-virus relationships | ||||
Official Date: | September 2012 | ||||
Dates: |
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Institution: | University of Warwick | ||||
Theses Department: | School of Life Sciences | ||||
Thesis Type: | PhD | ||||
Publication Status: | Unpublished | ||||
Supervisor(s)/Advisor: | Evans, D. J. (David J.) | ||||
Sponsors: | Biotechnology and Biological Sciences Research Council (BBSRC) | ||||
Extent: | xv, 337 leaves : illustrations, charts. | ||||
Language: | eng |
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