Trex1 exonuclease degrades ssDNA to prevent chronic checkpoint activation and autoimmune disease.

Summary Trex1 is the major 3′ DNA exonuclease in mammalian cells, and mutations in the human TREX1 gene can cause Aicardi-Goutieres syndrome, characterized by perturbed immunity. Similarly, Trex1 −/− mice have an autoinflammatory phenotype; however, the mechanism of Trex1-deficient disease is unknown. We report that Trex1, ordinarily associated with the endoplasmic reticulum (ER), relocalizes to the S phase nucleus after γ irradiation or hydroxyurea treatment. Notably, Trex1-deficient cells show defective G1/S transition and chronic ATM-dependent checkpoint activation, even in the absence of exogenous stress, correlating with persistent single-stranded DNA molecules produced in S phase, which accumulate in the ER. Our data indicate that Trex1 acts on a single-stranded DNA polynucleotide species generated from processing of aberrant replication intermediates to attenuate DNA damage checkpoint signaling and prevent pathological immune activation.