Skip to main content
. Author manuscript; available in PMC: 2009 Jun 20.
Published in final edited form as: Virology. 2008 Apr 18;376(1):42–52. doi: 10.1016/j.virol.2008.03.007

FIGURE 5.

FIGURE 5

A. Phosphopeptide map of pp71. A compilation of the results obtained with MS/MS analysis of phosphorylation sites of the pp71 tegument protein. The total analyzed sequence is underlined, and the phosphorylated residues are indicated in red. The region corresponding to the Mid-Region (MR, aa 200–300) is boxed. B. Transcriptional activity of pp71 phosphorylation site mutants. HFF cells were transfected with pMIEP-LUC and pSI71HAX or the indicated mutants. Cells were harvested 48 hours post-transfection and assessed for luciferase activity. Data is the average and standard deviation of a minimum of three experiments and is expressed relative to the activity of the wild type (WT) pp71 protein. Symbol: * significantly different from wild type, p<0.05. C. Subcellular localization of pp71 phosphorylation site mutants. HFF cells were transfected with constructs expressing the indicated pp71 mutants, and processed for immunofluorescence and confocal microscopy 48 hours after transfection using antibodies to the HA epitope tag and nuclear ND10 domains. The pp71 proteins are shown as red fluorescence, and ND10 domains shown as green fluorescence. D. Western Blot analysis of pp71 phosphorylation site mutants. HeLa cells were transfected with constructs expressing eGFP (lane 1), pp71 (lane 2), pp71T223A (lane 3), or pp71T223D (lane 4). Cells were harvested at 48 hours post-transfection and the extracts subjected to western blot analysis using an antibody to the HA-epitope tag. Symbol: * non-specific band used as a loading control.