Volume 8 issue 11 November 2007
Growth control of Golgi phosphoinositides by reciprocal localization of Sac1 lipid phosphatase and Pik1 4-kinase
Frank Faulhammer, Suparna Kanjilal-Kolar, Andreas Knödler, Jennifer Lo, Yerim Lee, Gerlinde Konrad, and Peter Mayinger

Supplemental Figure 1: Quantification of GFP-Sac1p localization in wild-type, rer1Δ and rer1Δ dpm1ΔC-sec62 double mutant strains. (A) Quantification of GFP-Sac1p localization under different growth conditions. A CEN-based plasmid for expressing GFP-Sac1p under control of the endogenous SAC1 promoter (pGK27) was introduced into a sac1Δ strain (ATY202) (A), a rer1Δ strain (YSC1021) (B) and a rer1Δ dpm1ΔC-sec62 double mutant (PMY153) (C). Growth conditions as indicated in the Figure. Data are from at least three independently grown cultures with ~300 cells analyzed for each condition (+/-SE).

Figure 1 (.jpg)

Supplemental Figure 2: Analysis of Sac1 localization mutants. (A) Lack of colocalization with Alg9p-RFP and localization during starvation. Nomarski optics and fluorescence colocalization of Alg9p-RFP and GFP-Sac1p (pGK27), GFP-Sac1Δ588-620 (pFF21), and GFP-Sac1-102 (pFF25). The coding region for Alg9p-RFP was integrated into the ALG9 locus in a sac1Δ strain (PMY148). (B) Localization of Sac1 mutant proteins during starvation. Nomarski optics and fluorescence localization of GFP-Sac1p (pGK27), GFP-Sac1Δ611-620 (pFF22), and GFP-Sac1-102 (pFF25) expressed in a sac1Δ strain (ATY 202) during exponential growth (Exp) and after glucose starvation (-Glu, 30 min).

Figure 2 (.jpg)