Figure 2

Yeast proteins phosphorylated by Dbf2-Mob1. (A) Of the 25 proteins with the highest phosphorylation signal as shown in Table 4, 22 of these genes were TAP-tagged in the Open Biosystems TAP-tagged yeast library. The TAP-tagged proteins were immunoprecipitated with IgG sepharose beads from asynchronous cultures, fractionated on SDS-PAGE and immunoblotted with anti-TAP. Of the 22 strains, 5 did not have detectable protein expression, such as VHS2 as shown. (B) The TAP-tagged proteins expressed in (A) were treated with ^FHHDbf2-^H6Mob1^TM9 in the presence of [γ-³²P]ATP, fractionated on SDS-PAGE and detected by autoradiography. (C) The TAP-tagged proteins phosphorylated by ^FHHDbf2-^H6Mob1^TM9 in (B) were treated with either ^FHHDbf2-^H6Mob1^TM9 or the kinase inactive ^FHHDbf2(N305A)-^H6Mob1^TM9 in the presence of [γ-³²P]ATP, fractionated on SDS-PAGE and detected by autoradiography.