FIG 3.

HPV31 E7 interacts with Nbs1 through the Mre11 binding domain. (A) Schematic of Nbs1 constructs depicting relevant binding domains and phosphorylation sites. (B) NBS-ILB1 cells stably expressing pLXIN vector alone or the indicated Nbs1 mutants were transiently transfected with HA-HPV31 E7. Immunoprecipitations were performed using an antibody to Nbs1, followed by Western blotting using an antibody to HA. Western blot analysis was performed on the indicated input lysates using antibodies to HA, Nbs1, and GAPDH. (C) NBS-ILB1 cells stably expressing pLXIN vector alone, pLXIN-Nbs1, or a pLXIN-Nbs1 Mre11 binding mutant (Mre11 BM) were transiently transfected with HA-HPV31 E7. Immunoprecipitation of whole-cell lysates was performed using an antibody to Nbs1 followed by Western blotting with an antibody to HA and Mre11. Western blot analysis was performed on the indicated input lysates using antibodies to HA, Nbs1, Mre11, and GAPDH. IP, immunoprecipitation; W, immunoblotting. All results are representative of observations of three independent experiments.