Functional characterization of human CTC 1 mutations reveals novel mechanisms responsible for the pathogenesis of the telomere disease C oats plus

Summary C oats plus is a rare recessive disorder characterized by intracranial calcifications, hematological abnormalities, and retinal vascular defects. This disease results from mutations in CTC 1, a member of the CTC 1– STN 1– TEN 1 ( CST ) complex critical for telomere replication. Telomeres are specialized DNA /protein structures essential for the maintenance of genome stability. Several patients with C oats plus display critically shortened telomeres, suggesting that telomere dysfunction plays an important role in disease pathogenesis. These patients inherit CTC 1 mutations in a compound heterozygous manner, with one allele encoding a frameshift mutant and the other a missense mutant. How these mutations impact upon telomere function is unknown. We report here the first biochemical characterization of human CTC 1 mutations. We found that all CTC 1 frameshift mutations generated truncated or unstable protein products, none of which were able to form a complex with STN 1– TEN 1 on telomeres, resulting in progressive telomere shortening and formation of fused chromosomes. Missense mutations are able to form the CST complex at telomeres, but their expression levels are often repressed by the frameshift mutants. Our results also demonstrate for the first time that CTC 1 mutations promote telomere dysfunction by decreasing the stability of STN 1 to reduce its ability to interact with DNA P olα, thus highlighting a previously unknown mechanism to induce telomere dysfunction.