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Cited by 286

The DOI matches its Crossref record; 3 of the 3 references it cites were found in a registry or an index.

Reconstitution of human telomerase activity in vitro

Beattie, Tara L., Zhou, Wen, Robinson, Murray O., Harrington, Lea·1998·Current Biology
DOI 10.1016/s0960-9822(98)70067-3
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Abstract

Telomerase is a ribonucleoprotein enzyme complex that adds single-stranded telomere DNA to chromosome ends [1]. The RNA component of telomerase contains the template for telomeric DNA addition and is essential for activity [1,2]. Telomerase proteins have been identified in ciliates, yeast and mammals [3-12]. In Saccharomyces cerevisiae, the Est2 protein is homologous to the 123 kDa reverse transcriptase subunit of Euplotes telomerase, and is essential for telomerase activity [8]. In humans, telomerase activity is associated with the telomerase RNA hTR [13], the telomerase RNA-binding protein TP1/TLP1 [5,12] and the TP2 protein encoded by the human EST2 homolog [12] (also known as TRT1, hEST2 or TCS1 [9-11]). The minimal complex sufficient for activity is, however, unknown. We have reconstituted human telomerase activity in reticulocyte lysates and find that only exogenous hTR and TP2 are required for telomerase activity in vitro. Recognition of telomerase RNA by TP2 was species specific, and nucleotides 10-159 of hTR were sufficient for telomerase activity. Telomerase activity immunoprecipitated from the reticulocyte lysate contained hTR and recombinant TP2. Substitution of conserved amino acid residues in the reverse transcriptase domain of TP2 completely abolished telomerase activity. We suggest that TP2 and hTR might represent the minimal catalytic core of human telomerase.

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Research works printed but not linked: 21

  1. Greider, 1996, Telomere length regulation, Annu Rev Biochem, 65, 337, 10.1146/annurev.bi.65.070196.002005 · DOI
  2. Harrington, 1995, Gel shift and UV crosslinking analysis of Tetrahymena telomerase, J Biol Chem, 270, 8893, 10.1074/jbc.270.15.8893 · DOI
  3. Collins, 1995, Purification of Tetrahymena telomerase and cloning of genes encoding two protein components of the enzyme, Cell, 81, 677, 10.1016/0092-8674(95)90529-4 · DOI
  4. Harrington, 1997, A mammalian telomerase-associated protein, Science, 275, 973, 10.1126/science.275.5302.973 · DOI
  5. Nakayama, 1997, TLP1: a gene encoding a protein component of mammalian telomerase is a novel member of WD repeats family, Cell, 88, 875, 10.1016/S0092-8674(00)81933-9 · DOI
  6. Lingner, 1996, Purification of telomerase from Euplotes aediculatus: requirement of a primer 3′ overhang, Proc Natl Acad Sci USA, 93, 10712, 10.1073/pnas.93.20.10712 · DOI
  7. Lingner, 1997, Reverse transcriptase motifs in the catalytic subunit of telomerase, Science, 276, 561, 10.1126/science.276.5312.561 · DOI · Paper page
  8. Nakamura, 1997, Telomerase catalytic subunit homologs from fission yeast and human, Science, 277, 955, 10.1126/science.277.5328.955 · DOI · Paper page
  9. Meyerson, 1997, hEST2, the putative human telomerase catalytic subunit gene, is up-regulated in tumor cells and during immortalization, Cell, 90, 785, 10.1016/S0092-8674(00)80538-3 · DOI · Paper page
  10. Kilian, 1997, Isolation of a candidate human telomerase catalytic subunit gene, which reveals complex splicing patterns in different cell types, Hum Mol Genet, 6, 2011, 10.1093/hmg/6.12.2011 · DOI
  11. Harrington, 1997, Human telomerase contains evolutionarily conserved catalytic and structural subunits, Genes Dev, 23, 3109, 10.1101/gad.11.23.3109 · DOI · Paper page
  12. Autexier, 1996, Reconstitution of human telomerase activity and identification of a minimal functional region of the human telomerase RNA, EMBO J, 15, 5928, 10.1002/j.1460-2075.1996.tb00979.x · DOI
  13. Blasco, 1995, Functional characterization and developmental regulation of mouse telomerase RNA, Science, 269, 1267, 10.1126/science.7544492 · DOI
  14. Counter, 1997, The catalytic subunit of yeast telomerase, Proc Natl Acad Sci USA, 94, 9202, 10.1073/pnas.94.17.9202 · DOI
  15. Singer, 1994, TLC1: template RNA component of Saccharomyces cerevisiae telomerase, Science, 266, 404, 10.1126/science.7545955 · DOI · Paper page
  16. Lundblad, 1989, A mutant with a defect in telomere elongation leads to senescence in yeast, Cell, 57, 633, 10.1016/0092-8674(89)90132-3 · DOI
  17. Lendvay, 1996, Senescence mutants of Saccharomyces cerevisiae with a defect in telomere replication identify three additional EST genes, Genetics, 144, 1399, 10.1093/genetics/144.4.1399 · DOI
  18. Cohn, 1995, Telomerase in yeast, Science, 269, 396, 10.1126/science.7618104 · DOI
  19. Lingner, 1997, Three ever shorter telomere (EST) genes are dispensable for in vitro yeast telomerase activity, Proc Natl Acad Sci USA, 94, 11190, 10.1073/pnas.94.21.11190 · DOI
  20. Weinrich, 1997, Reconstitution of human telomerase with the template RNA component hTR and the catalytic protein subunit hTRT, Nat Genet, 17, 498, 10.1038/ng1297-498 · DOI · Paper page
  21. J Nakayama, H Tahara, E Tahara, M Saito, K Ito, H Nakamura, et al., Telomerase activation by hTRT in human normal fibroblasts and hepatocellular carcinomas, Nat Genet 18 65-68. · DOI · Paper page

Cited by

Metadata

Authors
Beattie, T. L., Zhou, W., Robinson, M. O., & Harrington, L.
Venue
Current Biology
Publisher
Elsevier BV
Published
1998-01
Volume
8
Issue
3
Pages
177-180
ISSN
0960-9822
DOI
10.1016/s0960-9822(98)70067-3
Cited by
286
References
3
Language
en
Licence

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