Chemical screening identifies ATM as a target for alleviating senescence

Abstract

Senescence, defined as irreversible cell-cycle arrest, is the main driving force of aging and age-related diseases. Here, we performed high-throughput screening to identify compounds that alleviate senescence and identified the ataxia telangiectasia mutated (ATM) inhibitor KU-60019 as an effective agent. To elucidate the mechanism underlying ATM's role in senescence, we performed a yeast two-hybrid screen and found that ATM interacted with the vacuolar ATPase V1 subunits ATP6V1E1 and ATP6V1G1. Specifically, ATM decreased E-G dimerization through direct phosphorylation of ATP6V1G1. Attenuation of ATM activity restored the dimerization, thus consequently facilitating assembly of the V1 and V0 domains with concomitant reacidification of the lysosome. In turn, this reacidification induced the functional recovery of the lysosome/autophagy system and was coupled with mitochondrial functional recovery and metabolic reprogramming. Together, our data reveal a new mechanism through which senescence is controlled by the lysosomal–mitochondrial axis, whose function is modulated by the fine-tuning of ATM activity.

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Figure 1: ATM as a potential target for ameliorating senescence.
Figure 2: ATM interacts with ATP6V1E1 and ATP6V1G1.
Figure 3: ATM controls V1-V0 assembly in the V-ATPase.
Figure 4: Recovery of mitochondrial function through activation of the lysosome/autophagy system.
Figure 5: Decreased ROS levels and abnormal nuclear morphology after KU-60019 treatment.
Figure 6: In vivo effects of KU-60019 on wound healing in aged mice.

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Acknowledgements

This research was supported by the Samsung Advanced Institute of Technology and the DGIST R&D Program of the Ministry of Science, ICT and Technology of Korea (20160165 to Y.-S.L. and 20160172 to S.C.P.).

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Contributions

H.T.K., J.T.P., and S.C.P. conceived and designed the experiments; H.T.K. elucidated lysosomal pH regulation by ATM and mitochondrial metabolic reprogramming by ATM inhibitor, and performed experiments including in vitro phosphorylation, confocal imaging, and flow cytometry detecting lysosomes, mitochondria, and autophagic flux; J.T.P. elucidated the effects of ATM inhibitor by HTS and performed wound-healing experiments and flow cytometry detecting lysosomes and mitochondria; K.C. elucidated the interactions between ATM and the V-ATPase V1 subunits, and performed subcellular fractionation; Y.K. performed site-directed mutagenesis; H.J.C.C. measured the frequency of abnormal nuclear morphology and performed neutral comet assays; C.W.J. assisted with HTS and wound-healing experiments; Y.-S.L. performed experiments with ATM shRNA and ATR inhibitor; H.T.K. and J.T.P. analyzed the data; J.T.P. supervised all experiments; J.T.P., H.T.K., Y.-S.L., and S.C.P. wrote and edited the paper.

Corresponding authors

Correspondence to Joon Tae Park or Young-Sam Lee or Sang Chul Park.

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The authors declare no competing financial interests.

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Kang, H., Park, J., Choi, K. et al. Chemical screening identifies ATM as a target for alleviating senescence. Nat Chem Biol 13, 616–623 (2017). https://doi.org/10.1038/nchembio.2342

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