| Summary: | The overall aim of this study was to characterise A1HV-1 A9. The specific aims were to determine: if the A9 gene product is functional; when in the viral life cycle A9 is expressed; where in the cell the protein is localised; whether any cellular or viral binding partners can be defined; is the A9 gene expressed in large granular lymphocyte (LGL) T cell lines? Northern blot analyses showed that A9 mRNA was expressed as an early gene in the productive phase of the virus life cycle. The A9 gene product was also shown to protect CHO cells from cis-platin induced apoptosis. The protein encoded by the A9 gene was successfully expressed in <i>E. coli</i> and was used to immunise mice and rabbits. Antibodies specific for the A9 protein were generated. However, using these antibodies, the localisation of A9 in CHO cells was inconclusive. A9 mRNA was expressed in LGL T cells from MCF-affected rabbits. To determine binding partners, an expression system involving tandem affinity purification (TAP) technique was used. Various cDNAs related to the bcl-2 family of proteins were successfully transfected into CHO cells as shown by RT-PCR and immunofluorescence analysis. However, MADLI/ mass spectrometry analysis of samples failed to identify specific binding partners for A9. A9 vbcl-2 is anti-apoptotic and may function to allow viral replication in infected cells in the productive phase of the life cycle.
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