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Figure 6


Fig. 6. The srv2-201 allele suppresses defects in cell growth and actin organization caused by pfy1-4. (A) Comparison of cell growth defects for progeny from a cross between pfy1-4 and srv2-201 strains. Diploids from this cross were sporulated, tetrads were dissected. Lethality of spores was scored on the tetrad plates. Viable haploid strains from these plates were re-plated on YPD and scored for cell growth at 25 and 37°C. n, number of haploid progeny with the indicated genotype analyzed. (B) Transformation assay showing that srv2-201 suppresses the growth defects of pfy1-4. Double mutant pfy1-4, srv2-201 cells were transformed with a low copy URA-marked empty vector (pRS316) or pSRV2 (pBG334). Serial dilutions of transformed cells were plated on Ura- selective medium and grown at 25, 30, 34 and 37°C. As a control, double mutant aip1, srv2-201 cells were transformed with same vectors. (C) Cells from A above were grown to log phase, chemically fixed and stained with Alexa Fluor 488-phalloidin to label F-actin structures. (D) Quantitative comparison of actin patch polarization defects in pfy-4 and pfy1-4 srv2-201 cells. Medium-budded cells (n>200, each strain) were scored for actin patch distribution in the bud. Actin patches were classified as being polarized, unpolarized or intermediate polarized.





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