title: Senescence mutants of Saccharomyces cerevisiae with a defect in telomere replication identify three additional EST genes.
abstract: The primary determinant for telomere replication is the enzyme telomerase, responsible for elongating the G-rich strand of the telomere. The only component of this enzyme that has been identified in Saccharomyces cerevisiae is the TLC1 gene, encoding the telomerase RNA subunit. However, a yeast strain defective for the EST1 gene exhibits the same phenotypes (progressively shorter telomeres and a senescence phenotype) as a strain deleted for TLC1, suggesting that EST1 encodes either a component of telomerase or some other factor essential for telomerase function. We designed a multitiered screen that led to the isolation of 22 mutants that display the same phenotypes as est1 and tlc1 mutant strains. These mutations mapped to four complementation groups: the previously identified EST1 gene and three additional genes, called EST2, EST3 and EST4. Cloning of the EST2 gene demonstrated that it encodes a large, extremely basic novel protein with no motifs that provide clues as to function. Epistasis analysis indicated that the four EST genes function in the same pathway for telomere replication as defined by the TLC1 gene, suggesting that the EST genes encode either components of telomerase or factors that positively regulate telomerase activity.
authors.affiliation: Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.
authors.forename: T S
authors.initials: TS
authors.lastname: Lendvay
authors.forename: D K
authors.initials: DK
authors.lastname: Morris
authors.forename: J
authors.initials: J
authors.lastname: Sah
authors.forename: B
authors.initials: B
authors.lastname: Balasubramanian
authors.forename: V
authors.initials: V
authors.lastname: Lundblad
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004271:DNA, Fungal / Q000235:genetics; D004268:DNA-Binding Proteins; D005656:Fungal Proteins / Q000235:genetics*; D015966:Gene Expression Regulation, Fungal; D008969:Molecular Sequence Data; D009154:Mutation; D011506:Proteins / Q000235:genetics; D012313:RNA; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D029701:Saccharomyces cerevisiae Proteins; D019098:Telomerase / Q000235:genetics*; D016615:Telomere / Q000235:genetics*; D034501:Telomere-Binding Proteins
chemical_list: C452219:Cdc13 protein, S cerevisiae; D004271:DNA, Fungal; D004268:DNA-Binding Proteins; D005656:Fungal Proteins; D011506:Proteins; D029701:Saccharomyces cerevisiae Proteins; D034501:Telomere-Binding Proteins; C422952:telomerase RNA; D012313:RNA; C099024:EST1 protein, S cerevisiae; C509185:EST2 protein, S cerevisiae; C491580:EST3 protein, S cerevisiae; D019098:Telomerase