Transient telomerase inhibition alters cell cycle kinetics

Abstract

Telomerase is the ribonucleoprotein reverse transcriptase that catalyzes the synthesis of telomeres at the ends of linear human chromosomes and contributes to proper telomere-loop (T-loop) formation. Formation of the T-loop, an obligate step before cell division can proceed, requires the generation of a 3-overhang on the G-rich strand of telomeric DNA via telomerase or C-strand specific nucleases. Here, we discover telomerase activity is critical for efficient cell cycle progression using transient chemical inhibition by the telomerase inhibitor imetelstat. Telomerase inhibition caused changes in cell cycle kinetics and increased the proportion of cells in G2 phase, suggesting delayed clearance through this checkpoint. Investigating the possible contribution of unstructured telomere ends to these cell cycle distribution changes, we observed that imetelstat treatment induced {gamma}H2AX DNA damage foci in a subset of telomerase-positive cells but not telomerase-negative primary human fibroblasts. Chromatin-immunoprecipitation with {gamma}H2AX antibodies demonstrated imetelstat treatment-dependent enrichment of this DNA damage marker at telomeres. Notably, the effects of telomerase inhibition on cell cycle profile alterations were abrogated by pharmacological inhibition of the DNA-damage-repair transducer ATM.\n\nAdditionally, imetelstat potentiation of etoposide, a DNA-damaging drug that acts preferentially during S/G2 phases of the cell cycle, also depended on functional ATM signaling. Our results suggest that telomerase inhibition delays the removal of ATM-dependent DNA damage signals from telomeres in telomerase-positive cancer cells. This demonstrates for the first time that telomerase activity directly facilitates the progression of the cell cycle through modulation of transient telomere dysfunction signals.