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. 1999 Oct;10(10):3331–3343. doi: 10.1091/mbc.10.10.3331

Figure 2.

Figure 2

UV and bleomycin sensitivities and thermosensitive colony formation of rad22 and rti1 mutants. (A) UV and bleomycin sensitivities of Δrti1 and Δrad22 cells. Wild-type (ATCC38399; square), rad1-1 (HM73; cross), Δrad22 (HM367; circle), and Δrti1 (HM368; closed diamond) cells were plated on MMA + leu plates at 500 cells per plate, irradiated with various doses of short UV, and incubated at 23°C for 7 d. Sensitivity to bleomycin was determined by plating on MMA + leu plates containing the indicated concentrations of bleomycin. Percent cell viability was calculated by dividing the number of formed colonies by the number of plated cells. (B) Temperature-sensitive colony formation of rad22 and rti1 mutants. Wild-type (K150-A13; square), Δrad22 (HM367; circle), Δrti1 (HM368; closed diamond), rad22-H6 (HM366; triangle), and rad22-H6 Δrti1 (HM369; square with plus) cells were grown in YEA at 23°C for 1 d, transferred in YE medium at 23°C, and incubated for 14 h. One thousand cells were plated onto YEA plates and incubated at the temperatures specified for 7 d. Percent colony formation was calculated by dividing the number of formed colonies by the number of plated cells. (C) rad22+/rad22+ diploid (KS-1; square), rad22+/rad22-H6 diploid (KS-2; diamond), and rad22-H6/rad22-H6 diploid (KS-3; circle) cells were grown in PM + leu at 23°C for 16 h. Five hundred cells of each strain were plated on to MMA + leu plates and incubated at the indicated temperatures for 5 d. Percent colony formation was calculated by dividing the number of formed colonies by the number of plated cells. (D) Tetrad analysis of Δrad22 Δrti1 double disruptants. The h− rad22::ura4+ leu1-32 ura4-D18 cells (HM375) and the h+ rti1::ura4+ leu1-32 ura4-D18 cells (HM3368) were mixed and plated on a MEA plate and incubated at 25°C for 3 d for conjugation, meiosis, and sporulation. Spores were then isolated from each ascus and placed on YEA followed by incubation at 25°C for 5 d. (E) Morphology of geminated Δrad22 Δrti1 double disruptant cells. Double disruptant spores were germinated at 25°C for 5 d and photographed. Germinated wild-type cells in the same experiment were suspended in YE and photographed for comparison of cell size. (F) Tetrad analysis of rad22-H6 mat1-p Δ17 double mutants. The h− rad22-H6 leu1-32 (AN1) cells were crossed to h+ mat1-p Δ17 leu1-32 ura4-D18 cells. Formed spore asci were tetrad dissected, grown on YEA plates at 25°C, and examined for the ability to grow at 36.5°C and mating type. The mating type was determined by crossing to h− leu1-32 or h+ leu1-32 cells.