Abstract
Telomere dysfunction is typically studied under conditions in which a component of the six-subunit shelterin complex that protects chromosome ends is disrupted. The nature of spontaneous telomere dysfunction is less well understood. Here we report that immortalized human cell lines lacking wild-type p53 function spontaneously show many telomeres with a DNA damage response (DDR), commonly affecting only one sister chromatid and not associated with increased chromosome end-joining. DDR+ telomeres represent an intermediate configuration between the fully capped and uncapped (fusogenic) states. In telomerase activity–positive (TA+) cells, DDR is associated with low TA and short telomeres. In cells using the alternative lengthening of telomeres mechanism (ALT+), DDR is partly independent of telomere length, mostly affects leading strand–replicated telomeres, and can be partly suppressed by TRF2 overexpression. In ALT+ (but not TA+) cells, DDR+ telomeres preferentially associate with large foci of extrachromosomal telomeric DNA and recombination proteins. DDR+ telomeres therefore arise through different mechanisms in TA+ and ALT+ cells and have different consequences.
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Acknowledgements
We thank T. de Lange for supplying the TRF2ΔBΔM construct used in these experiments, and members of the Children's Medical Research Institute for comments on the manuscript. This work was supported by a US National Science Foundation international research fellowship (0602009), a grant from the Cure Cancer Australia Foundation and a Sir Keith Murdoch fellowship from the American Australian Association (to A.J.C.); a Promina postdoctoral fellowship (to H.A.P.); an Australian Postgraduate Award and a Judith Hyam Memorial Trust Fund for Cancer Research scholarship (to Z.K.) and a Cancer Council New South Wales program grant (to R.R.R.).
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A.J.C., Z.K., S.B.C., C.E.N., H.A.P. and A.A.N. conducted the experiments. A.J.C, A.A.N. and R.R.R. designed the project. A.J.C. and R.R.R. analyzed the data and authored the manuscript.
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Cesare, A., Kaul, Z., Cohen, S. et al. Spontaneous occurrence of telomeric DNA damage response in the absence of chromosome fusions. Nat Struct Mol Biol 16, 1244–1251 (2009). https://doi.org/10.1038/nsmb.1725
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DOI: https://doi.org/10.1038/nsmb.1725
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