sensitivity of yeast strains with long g-tails to levels of telomere-bound telomerase灵敏度的酵母菌株长g-tails telomere-bound端粒酶水平.pdfVIP

Sensitivity of Yeast Strains with Long G-Tails to Levels of Telomere-Bound Telomerase 1[¤ 1[ 2 2 1 Leticia R. Vega , Jane A. Phillips , Brian R. Thornton , Jennifer A. Benanti , Mutiat T. Onigbanjo , 2 1* David P. Toczyski , Virginia A. Zakian 1 Department of Molecular Biology, Princeton University, Princeton, New Jersey, United States of America, 2 Cancer Research Institute, Department of Biochemistry and Biophysics, University of California, San Francisco California, United States of America The Saccharomyces cerevisiae Pif1p helicase is a negative regulator of telomere length that acts by removing telomerase from chromosome ends. The catalytic subunit of yeast telomerase, Est2p, is telomere associated throughout most of the cell cycle, with peaks of association in both G1 phase (when telomerase is not active) and late S/G2 phase (when telomerase is active). The G1 association of Est2p requires a specific interaction between Ku and telomerase RNA. In mutants lacking this interaction, telomeres were longer in the absence of Pif1p than in the presence of wild-type PIF1, indicating that endogenous Pif1p inhibits the active S/G2 form of telomerase. Pif1p abundance was cell cycle regulated, low in G1 and early S phase and peaking late in the cell cycle. Low Pif1p abundance in G1 phase was anaphase-promoting complex dependent. Thus, endogenous Pif1p is unlikely to act on G1 bound Est2p. Overexpression of Pif1p from a non-cell cycle-regulated promoter dramatically reduced viability in five strains with impaired end protection (cdc13–1, yku80D, yku70D, yku80–1, and yku80–4), all of which have longer single- strand G-tails than wild-type