Telomere attrition and other forms of telomere damage can activate the ATM kinase pathway. What generates the DNA damage signal at mammalian chromosome ends or at other double-strand breaks is not known. Telomere dysfunction is often accompanied by disappearance of the 3′ telomeric overhang^,, raising the possibility that DNA degradation could generate the structure that signals. Here we address these issues by studying telomere structure after conditional deletion of mouse TRF2, the protective factor at telomeres. Upon removal of TRF2 from TRF2^F/− p53^−/− mouse embryo fibroblasts, a telomere damage response is observed at most chromosome ends. As expected, the telomeres lose the 3′ overhang and are processed by the non-homologous end-joining pathway. Non-homologous end joining of telomeres was abrogated in DNA ligase IV-deficient (Lig4^−/−) cells. Unexpectedly, the telomeres of TRF2^−/− Lig4^−/− p53^−/− cells persisted in a free state without undergoing detectable DNA degradation. Notably, the telomeres retained their 3′ overhangs, but they were recognized as sites of DNA damage, accumulating the DNA damage response factors 53BP1 and γ-H2AX, and activating the ATM kinase. Thus, activation of the ATM kinase pathway at chromosome ends does not require overhang degradation or other overt DNA processing.