Figure 6.

A Pan1 partial loss-of-function mutation partially suppresses the temperature sensitivity of the sla2Δ mutant. (A) The pan1-KE mutation partially suppresses the temperature sensitivity of the sla2Δ mutant at 34°C. A dilution series of cells was plated on YPD medium and incubated for the indicated times at 25, 34, or 37°C. (B) SDS-PAGE of a cosedimentation assay using full-length Pan1p, GST-CC, and F-actin. The gel was stained with Coomassie blue. P, pellets; S, supernatants. Indicated amounts of yeast actin were used with 100 nM Pan1p and 0, 250, or 500 nM GST-CC. (C) Diagram of wild-type Pan1p and Pan1ΔWA mutant. (D) Immunoblots showing that Pan1ΔWA is expressed at levels similar to wild-type Pan1p. Top panel was probed with anti-Pan1p antibody and bottom panel was probed with anti-Act1p antibody as a loading control. (E) Plates showing growth phenotypes of double-mutant combinations. Only pan1ΔWA results in a synthetic temperature sensitivity at 30°C when combined with arp2-1. All pan1 mutants result in synthetic lethality with arp2-1 on 3% formamide (FA) plates at 25°C. (F) The pan1ΔWA mutation partially suppresses the temperature sensitivity of the sla2Δ mutant at 34°C. A dilution series of cells was plated on YPD medium and incubated for indicated time at 25 or 34°C, respectively.