Wilcock, Diane; (1992) Effects of DNA tumour virus infection on host nuclear proteins. Doctoral thesis (Ph.D), UCL (University College London).

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Abstract

Immunofluorescence techniques have been used to investigate the distributions of a number of host cell proteins upon infection with three different DNA tumour viruses: herpes simplex virus type 1 (HSV-1), adenovirus (types 2, 4 and 5) and simian virus 40 (SV40). A number of host DNA replication proteins were shown to specifically redistribute to viral replication "compartments" labelled by antibody to the HSV-1 major DNA-binding protein (ICP8) during productive HSV-1 infection. These host proteins also colocalised to varying extents with ICP8 in the presence of a specific inhibitor of viral DNA replication which caused a marked alteration in the location of intranuclear ICPB. Two anti- oncogenic proteins, retinoblastoma and p53 were shown to relocate in the same manner as the cellular DNA replication proteins during both productive and abortive HSV-1 infections, raising the possibility that these proteins may be associated with DNA replication complexes in uninfected cells. Transfection experiments showed that the seven HSV-1 proteins shown to be essential for DNA replication can induce replication compartment formation in uninfected cells. Host DNA replication proteins were also redistributed during productive (but not abortive) infection with adenovirus, although, of those tested, only proliferating cell nuclear antigen (PCNA) was detected in viral replication compartments. A specific population of PCNA was also detected at novel discrete nuclear foci. SV40 infection resulted in the redistribution of singlestranded DNA-binding protein (SSB) to foci which enlarged as infection progressed and also contained SV40 large T antigen. SV40 infection of adenovirus-transformed 293 cells caused p53 to move from the cytoplasm to the nucleus. SV40 co-infections with adenovirus and HSV-1 were also studied. The production and characterisation of monoclonal antibodies to the HSV-1 major DNA binding protein (ICP8) and polymerase accessory factor (UL42) are also described.

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