FIG. 7.
Intracellular distribution of viral proteins is disrupted in L47A mutant-infected cells. Cells were infected with either wild-type (Wt) or L47A mutant (L47A) virus, separated into NP-40 cytoplasmic (Cytoplasm), and nuclear (Nucleus) fractions on days 7, 12, or 14 after infection and subjected to Western immunoassay using either anti-MCP (A) or anti-pAPhr (B), all as described in Results and Materials and Methods. Phosphorimages of the two resulting membranes are shown in panels A and B. (C) Calculations based on measurements from these membranes show the relative percentages of proteins of interest in the NP-40 cytoplasmic and nuclear fractions. Measurements for L47A from panel A included the MCP and “Frag” bands for all six samples and also the two smaller fragments (see asterisks) in “Nucleus” samples from days 12 and 14. Abbreviations indicate the 118-kDa fragment of MCP (Frag.), smaller fragments of MCP (*), protease precursor (pPR), protease precursor cleaved at M site and lacking carboxyl “tail” (PR, §), carboxyl fragment of pPR resulting from R-site cleavage (pPRc, linker+scaffold in Fig. 1), pPRc after removal of the carboxyl “tail” by M-site cleavage (−), assembly protein precursor (pAP), precursor of pUL80.4 (⋄), product of pAP cleavage at M site (AP), and precursor of pUL80.3 (p80.3).