Table 1.

Analysis of ICP8 alanine substitution mutants

ICP8 mutant	Protein expression	Protein localization	% Complementation
ICP8 WT	Yes	Nuclear	100
Y20A–F61A–Y90A	Yes	Cytoplasmic	0
C116A–R120A	Yes	Nuclear	70
R262A–H266A	Yes	Nuclear	14
R644A–D645A	Yes	Nuclear	54
Q706A–F707A	Yes	Nuclear	0
N882A–L883A	Yes	Nuclear	14
D905A–Y909A	No	N/A	N/A

Table showing the analysis of alanine substitution mutants assessed in this study. Vero cells were transiently transfected with 50 ng of mammalian expression plasmids containing ether WT or mutant versions of ICP8 and analyzed by Western blot and IF with anti-ICP8 antibody for protein expression and localization, respectively. For complementation analysis, transfected Vero cells were superinfected with HD-2 at an MOI of 3 for 24 h. Virus was harvested and titrated on the ICP8-complementing cell line (S2). Viral plaques were counted and reported as a percentage of WT ICP8. Results for Q706A and F707A mutant are in bold.