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-H6Mob1TM9 in the presence of [γ-32P]ATP, fractionated on SDS-PAGE and detected by autoradiography. (C) The TAP-tagged proteins phosphorylated by FHHDbf2-H6Mob1TM9 in (B) were treated with either FHHDbf2-H6Mob1TM9 or the kinase inactive FHHDbf2(N305A)-H6Mob1TM9 in the presence of [γ-32P]ATP, fractionated on SDS-PAGE and detected by autoradiography.
Mah et al. BMC Biochemistry 2005 6:22 doi:10.1186/1471-2091-6-22