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Yeast telomerase is capable of limited repeat addition processivity.

TitleYeast telomerase is capable of limited repeat addition processivity.
Publication TypeJournal Article
Year of Publication2004
AuthorsBosoy D, Lue NF
JournalNucleic Acids Res
Volume32
Issue1
Pagination93-101
Date Published2004
ISSN1362-4962
KeywordsBase Sequence, Deoxyguanine Nucleotides, DNA, DNA Primers, DNA Replication, Enzyme Stability, Mutation, RNA, Saccharomyces cerevisiae, Telomerase, Templates, Genetic
Abstract

Telomerase is a ribonucleoprotein reverse transcriptase responsible for the maintenance of one strand of telomere terminal repeats. Telomerase-mediated sequence addition is dictated by a short 'template' region of the RNA component. Despite the short template segment, telomerases from many organisms have been shown to mediate the synthesis of long extension products. This synthesis presumably depends on two types of translocation events: simultaneous translocation of the RNA-DNA duplex relative to the active site after each nucleotide incorporation (type I or nucleotide addition processivity), and translocation of the RNA relative to the DNA product after each round of repeat synthesis (type II or repeat addition processivity). In contrast, telomerases from yeasts have been shown to synthesize mostly short products, implying a defect in one or both types of translocation. In this report, we analyzed the processivity of yeast telomerase in vitro, and identified two position-specific elongation barriers within the 5' region of the RNA template that can account for the synthesis of incomplete first round products. These barriers respond differently to variations in nucleotide concentration, primer sequence and mutations in the catalytic protein subunit, consistent with their having distinct mechanistic bases. In addition, by using optimal primers and high concentrations of dGTP, we were able to detect significant type II translocation by the yeast enzyme. Thus, the difference between the elongation property of yeast and other telomerases appears to be quantitative rather than qualitative. Our results suggest that yeast may be a useful system for investigating the physiologic significance of repeat addition processivity.

DOI10.1093/nar/gkg943
Alternate JournalNucleic Acids Res
PubMed ID14704347
PubMed Central IDPMC373262

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