Direct involvement of the TEN domain at the active site of human telomerase

Julie Jurczyluk, Amanda S Nouwens, Jessica K. Holien, Timothy E Adams, George O. Lovrecz, Michael W. Parker, Scott B. Cohen, Tracy M. Bryan

Research output: Contribution to journalArticleResearchpeer-review

37 Citations (Scopus)

Abstract

Telomerase is a ribonucleoprotein that adds DNA to the ends of chromosomes. The catalytic protein subunit of telomerase (TERT) contains an N-terminal domain (TEN) that is important for activity and processivity. Here we describe a mutation in the TEN domain of human TERT that results in a greatly increased primer Kd, supporting a role for the TEN domain in DNA affinity. Measurement of enzyme kinetic parameters has revealed that this mutant enzyme is also defective in dNTP polymerization, particularly while copying position 51 of the RNA template. The catalytic defect is independent of the presence of binding interactions at the 5′-region of the DNA primer, and is not a defect in translocation rate. These data suggest that the TEN domain is involved in conformational changes required to position the 3′-end of the primer in the active site during nucleotide addition, a function which is distinct from the role of the TEN domain in providing DNA binding affinity.

Original languageEnglish
Pages (from-to)1774-1788
Number of pages15
JournalNucleic Acids Research
Volume39
Issue number5
DOIs
Publication statusPublished - Mar 2011
Externally publishedYes

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