Tel1ATM and Rad3ATR kinases promote Ccq1-Est1 interaction to maintain telomeres in fission yeast

Abstract

SUMMARYThe shelterin complex plays both positive and negative roles in telomerase regulation. While shelterin prevents the checkpoint kinases ATM and ATR from fully activating DNA damage responses at telomeres, those kinases are also required for telomere maintenance. In fission yeast, cells lacking both Tel1 (ATM ortholog) and Rad3 (ATR ortholog) fail to recruit telomerase to telomeres, and survive by circularizing chromosomes. However, the critical telomere substrate(s) of Tel1ATM/Rad3ATR remained unknown. Here, we show that Tel1ATM/Rad3ATR-dependent phosphorylation of the shelterin subunit Ccq1 on Thr93 is essential for telomerase association with telomeres. In addition, we show that the telomerase subunit Est1 interacts directly with the phosphorylated Thr93 of Ccq1 to ensure telomere maintenance. The shelterin subunits Taz1, Rap1 and Poz1 (previously established inhibitors of telomerase) were also found to negatively regulate Ccq1 phosphorylation. These findings establish Tel1ATM/Rad3ATR-dependent Ccq1 Thr93 phosphorylation as a critical regulator of telomere maintenance in fission yeast.