Control of telomerase recruitment and end protection by independent shelterin components

Telomeres are proposed to alternate between “closed” states, in which chromosome ends are protected from DNA damage signaling and inaccessible to telomerase, and “open” states, where they become accessible for telomerase mediated elongation but less protected. Whether these states reflect distinct molecular mechanisms or mutually exclusive structural conformations remains unclear. Here, we develop a single-cell assay to monitor telomerase activity in mouse embryonic stem cells. Using this approach, we demonstrate that the shelterin component TPP1 is essential for telomerase recruitment via its interaction with TIN2, independently of POT1. In contrast, POT1 is dispensable for telomerase function but required for telomere end protection, acting independently of TPP1. These findings challenge the classical open-closed telomere model and reveal that telomerase recruitment and end protection are mediated by genetically and molecularly separable mechanisms. Overall design: We sequenced genomic DNA enriched for sequences that are not cut by frequence cutters to detect incorpration of mutant repeats at telomeres