The Herpesvirus group has a front rank reputation in the field of herpesvirus molecular biology and is particularly active in the area of transcriptional regulation by VP16 and HCF function and interactions. More recently the group has expanded into areas of virus maturation including investigation of nuclear membrane reorganisation, and tegument protein compartmentalisation, adding to the original construction of viruses containing GFP fusion proteins with the successful construction of a recombinant virus expressing GFP linked to VP16. The group is also involved in the application of the unusual trafficking properties of the virus protein VP22 in the field of gene and protein delivery.

Current research projects include:

Nuclear Envelope Reorganisation during HSV infection. This work focuses on how HSV matures through the inner nuclear envelope, what changes there are to the underlying lamina and inner nuclear envelop membrane proteins, and which virus proteins are responsible. In particular current studies involve analysis of the reorganisation of e.g. lamin B receptor-GFP fusion proteins, or the lamins themselves, at various stages after infection in live cells, and biochemical aspects of the distribution of lamin subspecies into the soluble phase .

Above: Co-localisation between viral protein gB and lamin B receptor-GFP fusion in an infected cell

Above: NMR structure of SUM0-1

The Role of the SUMO Pathway in Subnuclear Targeting and its Disruption by the HSV RING Finger Protein ICP0. ICP0 (IE110k) is a RING finger protein involved in broad spectrum activation of all phases of HSV gene expression and appears to function by disrupting the normal modification, trafficking and stability of a class of host cell proteins. This project focuses on the links between SUMO (a small ubiquitin-like protein) modification and sub-nuclear compartmentalisation, how ICP0 disrupts this and the detailed mechanism of how this ultimately leads to global activation of gene expression.

VP16 and HCF trafficking. The basic molecular biology and biochemistry of the mechanism of action of VP16 is now understood due to the results of the host laboratory together with several other groups. Among the gaps in our knowledge is the role of HCF and its normal cellular function(s), how and to where VP16 is targeted in the cell (including neurons), how this changes with infection and how tegument proteins including VP16 and VP22 are recruited into the virion. This work has become the subject of a main programme of investigation, now waiting to be supplemented by the recruitment of the applicant. In addition with regard to VP16 trafficking in infected cells, a recombinant virus expressing VP16 linked to the Green Fluorescent Protein has been successfully constructed and compartmentalisation of VP16-GFP in living cells is currently under investigation.

Above: Stable cell line with GFP-HCF in the nucleus

Above: VP22 protein delivery to cells

VP22 as a novel delivery agent. This work is aimed at the development of VP22 as a novel gene and protein delivery agent. The prospect is to amplify the number of cells containing the candidate gene by virtue of the reported trafficking properties of VP22. Current work is expanding on these previous applications in gene therapy, and is focused on exploiting the VP22 protein itself novel delivery agent for small nucleic acids including antisense and ribozymes. We have demonstrated the assembly of a novel type of particle termed a Vectosome, and followed uptake and delivery of protein and oligonucleotides using time lapse microscopy.