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Molecular characterization of inter-telomere and intra-telomere mutations in human ALT cells

Nature Genetics volume 30, pages 301–305 (2002)Cite this article

Abstract

Telomeres in most immortal cells1,2,3 are maintained by the enzyme telomerase4, allowing cells to divide indefinitely. Some telomerase-negative tumors and immortal cell lines maintain long heterogeneous telomeres by the ALT (alternative lengthening of telomeres) mechanism5,6; such tumors are expected to be resistant to anti-telomerase drug therapies. Occasionally telomerase-negative Saccharomyces cerevisiae mutants survive, and 10% of them (type II survivors) have unstable telomeres7,8. As in human ALT+ cells9, short telomeres in yeast type II survivors lengthen abruptly; in yeast, this is dependent on the recombination proteins Rad52p and Rad50p10. In human cells, ALT involves copying of sequence from a donor to a recipient telomere11. We have characterized for the first time a class of complex telomere mutations seen only in ALT+ cells. The mutant telomeres are defined by the replacement of the progenitor telomere at a discrete point (fusion point) with a different telomere repeat array. Among 19 characterized fusion points, one occurred within the first six repeats of the telomere, indicating that these recombination-like events can occur anywhere within the telomere. One mutant telomere may have been involved in a secondary recombination-like mutation event, suggesting that these mutations are sporadic but ongoing in ALT+ cells. We also identified simple intra-allelic mutations at high frequency, which evidently contribute to telomere instability in ALT+ cells.

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Figure 1: The principle of TVR–PCR, and comparison of IIICF/a2 ALT– and ALT+ telomere maps.
Figure 2: Example of a mutation in the 12qΔ telomere in a WI-38 ALT+ clone.
Figure 3: Alignment of the ten mutant 12qΔ telomere codes from WI-38 ALT+ clones with the progenitor allele.

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Acknowledgements

We thank A.J. Jeffreys and C. May for their valuable comments on the manuscript. The work was funded by a UK Medical Research Council component group grant (to N.J.R.) and funding (to R.R.) from the Carcinogenesis Fellowship of the New South Wales Cancer Council. H.A.P. was supported by a UK-MRC studentship.

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Authors and Affiliations

  1. Department of Genetics, University of Leicester, University Road, Leicester, LE1 7RH, UK

    Helen Varley, Hilda A. Pickett, Jennifer L. Foxon & Nicola J. Royle

  2. Cancer Research Unit, Children's Medical Research Institute, Westmead, Sydney, Australia

    Roger R. Reddel

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  1. Helen Varley
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Correspondence to Nicola J. Royle.

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Varley, H., Pickett, H., Foxon, J. et al. Molecular characterization of inter-telomere and intra-telomere mutations in human ALT cells. Nat Genet 30, 301–305 (2002). https://doi.org/10.1038/ng834

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