FIGURE 9.

Cells expressing dpb11-m1,m2,m3,m5 exhibit no GINS·Mcm2-7 interaction. dpb11-td cells expressing DPB11-WT or dpb11-m1,m2,m3,m5 under restrictive conditions were arrested with α-factor and then released into medium lacking α-factor for the indicated times. A and B, left panels, whole cell extracts were analyzed by Western blot for expression of the indicated proteins. A, right panel, cell extracts were immunoprecipitated with antibodies directed against Dpb11, followed by Western analysis for Mcm2 or Sld2. B, right panel, cell extracts were prepared to isolate loaded Mcm2-7 complex as described previously (34). Antibodies directed against Mcm2 were used for immunoprecipitation, followed by Western blot analysis of Mcm2, Cdc45, or GINS.