FIG 4.

Mapping of UL51 domains required for UL51 interaction with UL14. (A) Schematic diagram of the UL51 gene product (top line), with its putative α-helix domains (black boxes), and diagrams of GST-UL51, the GST-UL51 deletion mutants, and the GST-UL51:90-130LIY/AAA mutant. The level of binding of each GST fusion protein to UL14 determined in the pulldown experiments shown in panels B and C is shown at the right of each protein. (B and C) The GST fusion proteins shown in panel A were immobilized on glutathione-Sepharose beads and mock reacted or reacted with lysates of Vero cells that had been infected with wild-type HSV-1(F) at an MOI of 5 for 18 h. The beads were washed extensively and divided into two parts. One part was analyzed by immunoblotting with anti-UL14 antibody (top gels), and the other was electrophoretically analyzed in a denaturing gel and CBB stained (bottom gels). (D) Amount of UL14 pulled down by GST, GST-UL51:90-130, or GST-UL51:90-130LIY/AAA shown in the top of panel C relative to those of the GST fusion protein shown in the bottom of panel C. Each value is the mean ± standard error from three independent experiments and is expressed relative to the mean value of GST, which was normalized to 100. Asterisks indicate significant differences: *, P < 0.05 (by two-tailed Student t test).