The Mre11 Cellular Protein Is Modified by Conjugation of Both SUMO-1 and SUMO-2/3 during Adenovirus Infection
Figure 3
The Mre11 protein is modified by addition of both SUMO-1 and SUMO-2/3 during Ad5 infection. (a) HFF cells either mock-infected or infected with Ad5, the E1B 55 kDa insertion mutant A143, or E1B 55 kDa, E4 Orf6, or E4 Orf3 null mutants were harvested at different times postinfection (10, 16, 28, and 36 h p.i.), and lysates were analyzed by immunoprecipitation and Western blot. The immunoprecipitation of Mre11 was performed using anti-Mre11, resolved on 10% SDS-PAGE, and visualized by immunoblotting using anti-SUMO-2/3 or anti-SUMO-1 mAbs. Specific Mre11-SUMO-2/3 or -SUMO-1 bands are indicated by (*). Western blotting was performed using anti-Mre11 as input control. DBP and actin were included as infection control and internal loading control, respectively. (b) Quantification of total SUMO-Mre11 protein. Nuclear extracts of HFF cells either mock-infected or infected with Ad5, A143, or virus null mutants (E1B 55 kDa, E4 Orf6, or E4 Orf3) were obtained at two different times postinfection (16 and 28 h p.i.). Nuclear extracts were incubated with the anti-Mre11, and SUMO was detected by a colorimetric assay, as described in Section 2.
