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G-tail telomere HPA: simple measurement of human single-stranded telomeric overhangs

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Abstract

Accurate measurement of telomeric 3′-overhang (G-tail) lengths is essential for investigation of the biological effects of telomere dysfunction. G-tail telomere hybridization protection assay (Gt-telomere HPA) has the advantages of being simple to perform, accurate and highly sensitive for G tails as short as 20 nucleotides. Furthermore, Gt-telomere HPA is specific and quantitative for human G tails, and can be used to assay cell lysates as well as genomic DNA.

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Figure 1: Strategy, linearity, specificity and sensitivity of Gt-telomere HPA.
Figure 2: Mean G-tail lengths in cultured human cells.

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References

  1. Makarov, V.L., Hirose, Y. & Langmore, J.P. Cell 88, 657–666 (1997).

    Article  CAS  Google Scholar 

  2. Wright, W.E., Tesmer, V.M., Huffman, K.E., Levene, S.D. & Shay, J.W. Genes Dev. 11, 2801–2809 (1997).

    Article  CAS  Google Scholar 

  3. Blackburn, E.H. Nature 408, 53–56 (2000).

    Article  CAS  Google Scholar 

  4. de Lange, T. Nat. Rev. Mol. Cell Biol. 5, 323–329 (2004).

    Article  CAS  Google Scholar 

  5. Chai, W., Shay, J.W. & Wright, W.E. Mol. Cell. Biol. 25, 2158–2168 (2005).

    Article  CAS  Google Scholar 

  6. Cimino-Reale, G. et al. Nucleic Acids Res. 29, E35 (2001).

    Article  CAS  Google Scholar 

  7. Saldanha, S.N., Andrews, L.G. & Tollefsbol, T.O. Eur. J. Biochem. 270, 389–403 (2003).

    Article  CAS  Google Scholar 

  8. Nakamura, Y. et al. Clin. Chem. 45, 1718–1724 (1999).

    CAS  PubMed  Google Scholar 

  9. Hirose, M. et al. J. Cancer Res. Clin. Oncol. 123, 337–344 (1997).

    Article  CAS  Google Scholar 

  10. Stewart, S.A. et al. Nat. Genet. 33, 492–496 (2003).

    Article  CAS  Google Scholar 

  11. van Steensel, B., Smogorzewska, A. & de Lange, T. Cell 92, 401–413 (1998).

    Article  CAS  Google Scholar 

  12. McElligott, R. & Wellinger, R.J. EMBO J. 16, 3705–3714 (1997).

    Article  CAS  Google Scholar 

  13. Riha, K., McKnight, T.D., Fajkus, J., Vyskot, B. & Shippen, D.E. Plant J. 23, 633–641 (2000).

    Article  CAS  Google Scholar 

Download references

Acknowledgements

This work was funded in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Culture, Sports, Science and Technology, Japan. We thank T. de Lange for providing the pBabepuro-NmycTRF2ΔBΔM. We thank A.M. Sera for critical reading of the manuscript and for technical assistance. We thank A. Shimamoto for helpful discussion.

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Correspondence to Hidetoshi Tahara.

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Supplementary information

Supplementary Fig. 1

AE-labeled telomere HPA probes (29 mer) specifically detect mammalian telomere DNA 5′-TTAGGG-3′ sequences. (PDF 541 kb)

Supplementary Fig. 2

Time dependent increase of luminescence by T7 exonuclease treatment. (PDF 523 kb)

Supplementary Fig. 3

T7 exonuclease treatment of genomic DNA from SiHa cancer cell line generated G-tails with mean lengths of over 1,600 nt. (PDF 517 kb)

Supplementary Fig. 4

Structure of synthetic telomeric G-tail DNA and possible hybridization patterns of the telomere HPA probe. (PDF 622 kb)

Supplementary Fig. 5

Point mutations 6 bases away from the acridinium ester (AE) labeled position do not affect the HPA signals on telomeres. (PDF 662 kb)

Supplementary Fig. 6

Linearity of HPA for the detection of Alu DNA. (PDF 571 kb)

Supplementary Fig. 7

Estimated mean G-tail lengths in cultured cells using Gt-telomere HPA. (PDF 550 kb)

Supplementary Methods (PDF 54 kb)

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Tahara, H., Kusunoki, M., Yamanaka, Y. et al. G-tail telomere HPA: simple measurement of human single-stranded telomeric overhangs. Nat Methods 2, 829–831 (2005). https://doi.org/10.1038/nmeth797

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  • DOI: https://doi.org/10.1038/nmeth797

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