A novel specificity for the primer-template pairing requirement in Tetrahymena telomerase.

Telomerase is a specialized reverse transcriptase with a built-in RNA template. Base pairing between the templating domain of telomerase RNA and a telomeric DNA primer is normally a characteristic of elongation of telomeric DNA. Here we demonstrate the mechanism by which Tetrahymena telomerase bypasses a requirement for template-primer pairing in order to add telomeric DNA de novo to completely non-telomeric DNA primers. We show that this reaction initiates by copying the template residue at the 3' boundary of the telomerase RNA template sequence. Unexpectedly, as the RNA template moves through the telomerase catalytic center, the number of required potential Watson-Crick base pairs between RNA template and DNA primer increases from zero to five. We propose that this unprecedented position specificity of a base pairing potential requirement in a polymerase underlies the chromosome healing mechanism of telomerase, and reflects constraints inherent in an internal template.