SUMO Pathway Dependent Recruitment of Cellular Repressors to Herpes Simplex Virus Type 1 Genomes (original) (raw)
Figure 6
Expression and analysis of mutants of Sp100A and their recruitment to sites associated with HSV-1 genomes.
A. Map of Sp100A and mutations introduced into EYFP-Sp100A fusion proteins. The sequence targeted by the anti-Sp100 shRNA and the silent mutations that render the mRNA resistant is marked (shSp100). B. Western blot of wt EYFP-Sp100A in control and Sp100 depleted cells, with anti-Sp100 and anti-EGFP antibodies (upper and lower parts, respectively). The bands are as follows: a - double modification of EYFP-Sp100A; b - single SUMO modification of EYFP-Sp100A; c - double modification of endogenous Sp100; d - unmodified EYFP-Sp100A; e - single SUMO modification of endogenous Sp100A; f - unmodified endogenous Sp100A. C. Western blot of wt and mutant Sp100 proteins in control and Sp100 depleted cells. The mutant ΔSSIM deletes residues 289–327 encompassing both the K297 SUMO modification site and the SIM.