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. 2003 May;23(9):3031–3042. doi: 10.1128/MCB.23.9.3031-3042.2003

FIG. 8.

FIG. 8.

Ectopic expression of polα+ in the polαts13 mutant can restore the myc-Trt1 protein and telomere length to near-wild-type level. (A) Ectopic expression of polα+ increases the cellular myc-Trt1 protein in polαts13 cells. Prior to the experiment, the polαs13 myc-trt1+ + polα+ (polαts13 + polα+) transformants had been grown at 36°C to select for expression of polα+. The myc-trt1+ (polα+), polαts13 myc-trt1+ (polαts13), and polαs13 myc-trt1+ + polα+ (polαts13 + polα+) strains were grown at 25°C to a cell density of 6 × 106 cells per ml (lanes 1 through 3) and then shifted to 36°C and continued to culture for 6 h (lanes 4 through 6). (B) Ectopic expression of polα+ restores myc-Trt1 to the wild-type level. The myc-trt1+ (polα+) and polαs13 myc-trt1+ + polα+ (polαts13 + polα+) cells were grown at 36°C for 15 h, and cell extracts were analyzed as described for panel A. (C) Ectopic expression of polα+ reduces telomere length in the polαts13 mutant. DNA was isolated from the same cell extracts as in panels A and B for analysis of the telomere lengths by Southern blotting. Lanes: 1, KG2 (wild type); 2, myc-trt1+ (polα+) (25°C); 3, polαts13 myc-trt1+ (polαts13) (25°C); 4, polαs13 myc-trt1+ + polα+ (polαts13 + polα+) (first selected at 36°C and then shifted to 25°C); 5, myc-trt1+ (polα+) (36°C, 6 h); 6, polαts13 myc-trt1+ (polαts13) (36°C, 6 h); 7, polαs13 myc-trt1+ + polα+ (polαts13 + polα+) (36°C, 6 h); 8, myc-trt1+ (polα+) (36°C, 15 h); 9, polαs13 myc-trt1+ + polα+ (polαts13 + polα+) (36°C, 15 h).