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

Nature Methods volume 2, pages 829–831 (2005)Cite this article

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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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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Authors and Affiliations

  1. Department of Cellular and Molecular Biology, Division of Integrated Medical Science, Graduate School of Biomedical Sciences, Hiroshima University, Kasumi 1-2-3, Minami-ku, Hiroshima City, Hiroshima, 734-8551, Japan

    Hidetoshi Tahara, Mayuko Kusunoki, Yusuke Yamanaka, Shusaku Matsumura & Toshinori Ide

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  1. Hidetoshi Tahara
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  2. Mayuko Kusunoki
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  3. Yusuke Yamanaka
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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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