Research Paper Volume 8, Issue 11 pp 2827—2847

Tethering telomerase to telomeres increases genome instability and promotes chronological aging in yeast


Figure 1. Expression of CDC13-EST2 promotes yeast chronological aging. (A) Telomere length analysis by Southern blot. The cells that contain pRS316 (Vector) or pRS316-CDC13-EST2 plasmid were passaged, and the genomic DNA from cells of different streakouts (labeled on top) was subjected to Southern blot assay using a telomeric TG1-3 probe. (B) Flow chart of CLS assay. (C) Spot assay of CLS. Both the normal-telomere and overlong-telomere cells of streakouts 4th and 5th were used to perform semi-quantitative CLS assay. The time of cultures in the CLS assay was labeled on the left (e.g. D3 means day 3). (D) Telomere length of cells used in (C) at D3 and D38 was examined by Southern blot with telomeric TG1-3 probe. (E) Colony formation unit (CFU) assay of CLS. CLS of cells BY4742-pRS316 (streakout 8th, normal telomeres) and BY4742 -pRS316-CDC13-EST2 (streakout 8th, overlong telomeres) was quantitatively examined. Survival (viable colonies) values at the indicated days of culture were normalized to CLS D1. Values are the averages of 6-10 cultures ± SEM. * denotes p<0.05, ** p<0.01, *** p<0.001 and **** p<0.0001. (F) Telomere length of cells in (E) at CLS D1 and CLS D27 was examined by Southern blot.