This thesis concerns gene 176R from camelpox virus (CMLV) that encodes a protein known as viral schlafen (v-slfn). v-slfn has an N-terminal domain related to the p26 protein from baculovirus and a C-terminal domain related to mammalian schlafen proteins. A full length v-slfn is expressed by all sequenced orthopoxviruses except vaccinia virus (VACV) and variola virus. The baculovirus p26 proteins are poorly characterised, with no known function. In contrast, murine schlafen (m-slfn) proteins are upregulated in response to infection and the promoter for m-slfn2 has NF-κB and AP-1 binding sites. The prototypic slfn, m-slfn1, halts cellular proliferation by inhibition of cyclin D1 expression in vitro and both m-slfn1 and m-slfn8 reduce thymocyte proliferation in vivo. v-slfn is a predominantly cytoplasmic protein of 57 kDa that is expressed both early and late during CMLV infection. Expression of v-slfn reverses the growth arrest resulting from m-slfn1 expression, and this is a result of a reversal of the inhibition of cyclin D1 expression. This effect can be seen following overexpression of various transcription factors that upregulate cyclin D1 expression. Recombinant VACV expressing enhanced levels of v-slfn replicated and spread at a comparable rate to control viruses in vitro, but was less virulent than controls in the intranasal model of infection in vivo. A group of viruses based on VACV WR were constructed, which lack the gene fragments (B2R and B3R) corresponding to CMLV 176R. The undisrupted sequence for 176R was also re-inserted at this locus, resulting in a virus that expresses v-slfn from its natural promoter. In vitro characterisation showed no differences in replication or spread when compared to controls. Thus, v-slfn is an orthologue of mammalian slfn proteins, and may exert its effect by reversing their inhibition of cellular proliferation.
| Identifer | oai:union.ndltd.org:bl.uk/oai:ethos.bl.uk:513438 |
| Date | January 2009 |
| Creators | Goodbody, Rory Eric |
| Contributors | Smith, Geoffrey L. ; Gubser, Caroline |
| Publisher | Imperial College London |
| Source Sets | Ethos UK |
| Detected Language | English |
| Type | Electronic Thesis or Dissertation |
| Source | http://hdl.handle.net/10044/1/5378 |
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