A Mutant with a Defect in Telomere Elongation.pdf

Cell, Vol. 57, 633-643, May 19, 1989, Copyright ? 1989 by Cell Press A Mutant with a Defect in Telomere Elongation Leads to Senescence in Yeast Victoria Lundblad* and Jack W. Szostak Department of Genetics Harvard Medical School and Department of Molecular Biology Massachusetts General Hospital Boston, Massachusetts 02114 Summary We describe a general assay designed to detect mu- tants of yeast that are defect ive for any of several aspects of telomere function. Using this assay, we have isolated a mutant that displays a progress ive de- crease in telomere length as well as an increased fre- quency of c h r o m o s o m e loss. This mutation def ines a new gene, des ignated E S ~ (for ever shorter telo- meres). Null alleles of EST1 are not immediately invia- ble- instead, they have a s e n e s c e n c e phenotype , due to the gradual loss of s e q u e n c e s essent ia l for telo- mere function, leading to a progress ive d e c r e a s e in chromosomal stability and s u b s e q u e n t cell death. Introduction Telomeres are specialized structures found at the natural ends of eukaryotic linear chromosomes, and they are re- quired both for chromosome stability and to permit the complete replication of the termini of chromosomes (re- viewed in Blackburn and Szostak, 1984). DNA sequence analysis has shown that nuclear telomeres are strikingly similar to each other in that they all consist of a tandemly repeated simple sequence in which one strand is dis- tinctly G-rich and the other strand is correspondingly C-rich. These repeats are always found in one orientation, with the G-rich strand running 5 to 3 toward the DNA ter- minus in Saccharomyces cerevisiae the repeated se- quence is 01_3A (for a review, see Blackburn, 1984). The recent demonstration that telomeres from higher eukary- otes as diverse as the flowering plant Arabidopsis thaliana and humans are composed of similar simple repeated se- quences (Richards