FIGURE 1:

Identification of myo2-iq1,5, a novel myosin mutant that phenocopies a loss of Sro7 function. (A) Schematic representation of the type V motor, Myo2. (B) Minimum domain required for binding to Sro7. Radiolabeled fragments of the neck region of Myo2 were obtained by in vitro translation and bound to purified protASro7 on beads. (C) Strategy for generating point mutations in the IQ motif and effects of such mutations on binding to Sro7. Alanine substitutions made are shown in red. Charts show the results of binding assays between radiolabeled wild-type Myo2 (782–990) or Myo2 mutants (782–990) and protASro7 or control GST-Mlc1 on beads. Error bars represent the SD from three independent experiments. Binding of the IQ region of Myo1 (761–932) is shown on the extreme right of each chart as a comparison. (D) Three individual colonies of haploid yeast containing mutations in the IQ domain of Myo2 were picked into microtiter wells, transferred to YPD at the indicated temperatures and salt concentrations, and compared with haploid yeast containing mutations in sro7Δ, sro77Δ, and sro7Δ; sro77Δ.