Collins, K., Greider, C. W. (November 1995) Utilization of ribonucleotides and RNA primers by Tetrahymena telomerase. Embo J, 14 (21). pp. 5422-32. ISSN 0261-4189 (Print)
Abstract
Telomerase is a ribonucleoprotein (RNP) DNA polymerase involved in telomere synthesis. A short sequence within the telomerase RNA component provides a template for de novo addition of the G-rich strand of a telomeric simple sequence repeat onto chromosome termini. In vitro, telomerase can elongate single-stranded DNA primers processively: one primer can be extended by multiple rounds of template copying before product dissociation. Telomerase will incorporate dNTPs or ddNTPs and will elongate any G-rich, single-stranded primer DNA. In this report, we show that Tetrahymena telomerase was able to incorporate a ribonucleotide, rGTP, into product polynucleotide. Synthesis of the product [d(TT)r(GGGG)]n was processive, suggesting that the chimeric product remained associated with the enzyme both at the active site and at a second, previously characterized, template-independent product binding site. As predicted by this finding, RNA-containing oligonucleotides served as primers for elongation. More than 3 nt of RNA at a primer 3' end decreased the quantity of product synthesis but increased the affinity of the primer for telomerase. Thus, RNA-containing primers were effective as competitive inhibitors of DNA primer elongation by telomerase. These results support the possible evolutionary origin of telomerase as an RNA-dependent RNA polymerase.
| Item Type: | Paper |
|---|---|
| Uncontrolled Keywords: | Animals Binding, Competitive DNA Primers/genetics/metabolism RNA/genetics/metabolism Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S. Ribonucleotides/genetics/metabolism Telomerase/ metabolism Tetrahymena/ enzymology |
| Subjects: | bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification organism description > animal bioinformatics > genomics and proteomics > genetics & nucleic acid processing > protein structure, function, modification > protein types > enzymes > telomerase |
| CSHL Authors: | |
| Communities: | CSHL labs |
| Depositing User: | Jessica Koos |
| Date: | 1 November 1995 |
| Date Deposited: | 14 Aug 2014 15:57 |
| Last Modified: | 14 Aug 2014 15:57 |
| PMCID: | PMC394651 |
| URI: | https://repository.cshl.edu/id/eprint/30580 |
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