Abstract
Ataxia telangiectasia mutated (ATM), a critical DNA damage sensor, also possesses non-nuclear functions owing to its presence in extra-nuclear compartments, including peroxisomes, lysosomes, and mitochondria. ATM is frequently altered in several human cancers. Recently, we and others have shown that loss of ATM is associated with defective mitochondrial autophagy (mitophagy) in ataxia–telangiectasia (A–T) fibroblasts and B-cell lymphomas. Further, we reported that ATM protein but not ATM kinase activity is required for mitophagy. However, the mechanism of ATM kinase activation during ionophore-induced mitophagy is unknown. In the work reported here, using several ionophores in A–T and multiple T-cell and B-cell lymphoma cell lines, we show that ionophore-induced mitophagy triggers oxidative stress–induced ATMSer1981 phosphorylation through ROS activation, which is different from neocarzinostatin-induced activation of ATMSer1981, Smc1Ser966, and Kap1Ser824. We used A–T cells overexpressed with WT or S1981A (auto-phosphorylation dead) ATM plasmids and show that ATM is activated by ROS-induced oxidative stress emanating from ionophore-induced mitochondrial damage and mitophagy. The antioxidants N-acetylcysteine and glutathione significantly inhibited ROS production and ATMSer1981 phosphorylation but failed to inhibit mitophagy as determined by retroviral infection with mt-mKeima construct followed by lysosomal dual-excitation ratiometric pH measurements. Our data suggest that while ATM kinase does not participate in mitophagy, it is activated via elevated ROS.
Change history
22 May 2021
A Correction to this paper has been published: https://doi.org/10.1007/s11010-021-04183-5
References
Vyas S, Zaganjor E, Haigis MC (2016) Mitochondria and cancer. Cell 166:555–566. https://doi.org/10.1016/j.cell.2016.07.002
Sun N, Youle RJ, Finkel T (2016) The mitochondrial basis of aging. Mol Cell 61:654–666. https://doi.org/10.1016/j.molcel.2016.01.028
Moro L (2020) Mitochondria at the crossroads of physiology and pathology. J Clin Med. https://doi.org/10.3390/jcm9061971
Youle RJ, Narendra DP (2011) Mechanisms of mitophagy. Nat Rev Mol Cell Biol 12:9–14. https://doi.org/10.1038/nrm3028
Lazarou M, Sliter DA, Kane LA, Sarraf SA, Wang C, Burman JL, Sideris DP, Fogel AI, Youle RJ (2015) The ubiquitin kinase PINK1 recruits autophagy receptors to induce mitophagy. Nature 524:309–314. https://doi.org/10.1038/nature14893
Vara-Perez M, Felipe-Abrio B, Agostinis P (2019) Mitophagy in cancer: a tale of adaptation. Cells. https://doi.org/10.3390/cells8050493
Chourasia AH, Boland ML, Macleod KF (2015) Mitophagy and cancer. CancerMetab 3:4. https://doi.org/10.1186/s40170-015-0130-8
Boland ML, Chourasia AH, Macleod KF (2013) Mitochondrial dysfunction in cancer. Front Oncol 3:292. https://doi.org/10.3389/fonc.2013.00292
Zhang J, Tripathi DN, Jing J, Alexander A, Kim J, Powell RT, Dere R, Tait-Mulder J, Lee JH, Paull TT, Pandita RK, Charaka VK, Pandita TK, Kastan MB, Walker CL (2015) ATM functions at the peroxisome to induce pexophagy in response to ROS. Nat Cell Biol 17:1259–1269. https://doi.org/10.1038/ncb3230
Kang HT, Park JT, Choi K, Kim Y, Choi HJC, Jung CW, Lee YS, Park SC (2017) Chemical screening identifies ATM as a target for alleviating senescence. Nat ChemBiol 13:616–623. https://doi.org/10.1038/nchembio.2342
Valentin-Vega YA, Maclean KH, Tait-Mulder J, Milasta S, Steeves M, Dorsey FC, Cleveland JL, Green DR, Kastan MB (2012) Mitochondrial dysfunction in ataxi A–T elangiectasia. Blood 119:1490–1500. https://doi.org/10.1182/blood-2011-08-373639
Qi Y, Qiu Q, Gu X, Tian Y, Zhang Y (2016) ATM mediates spermidine-induced mitophagy via PINK1 and Parkin regulation in human fibroblasts. Sci Rep 6:24700. https://doi.org/10.1038/srep24700
Sarkar A, Stellrecht CM, Vangapandu HV, Ayres M, Kaipparettu BA, Park JH, Balakrishnan K, Burks JK, Pandita TK, Hittelman WN, Neelapu SS, Gandhi V (2020) Ataxia telangiectasia mutated interacts with Parkin and induces mitophagy independent of kinase activity. Evidence from mantle cell lymphoma. Haematologica. https://doi.org/10.3324/haematol.2019.234385
Sarkar A, Balakrishnan K, Chen J, Patel V, Neelapu SS, McMurray JS, Gandhi V (2016) Molecular evidence of Zn chelation of the procaspase activating compound B-PAC-1 in B cell lymphoma. Oncotarget 7:3461–3476. https://doi.org/10.18632/oncotarget.6505
Lee JH, Mand MR, Kao CH, Zhou Y, Ryu SW, Richards AL, Coon JJ, Paull TT (2018) ATM directs DNA damage responses and proteostasis via genetically separable pathways. Sci Signal. https://doi.org/10.1126/scisignal.aan5598
Shiloh Y (2003) ATM and related protein kinases: safeguarding genome integrity. Nat Rev Cancer 3:155–168. https://doi.org/10.1038/nrc1011
So S, Davis AJ, Chen DJ (2009) Autophosphorylation at serine 1981 stabilizes ATM at DNA damage sites. J Cell Biol 187:977–990. https://doi.org/10.1083/jcb.200906064
Katayama H, Kogure T, Mizushima N, Yoshimori T, Miyawaki A (2011) A sensitive and quantitative technique for detecting autophagic events based on lysosomal delivery. ChemBiol 18:1042–1052. https://doi.org/10.1016/j.chembiol.2011.05.013
Smith GC, d'Adda di Fagagna F, Lakin ND, Jackson SP (1999) Cleavage and inactivation of ATM during apoptosis. Mol Cell Biol 19:6076–6084. https://doi.org/10.1128/mcb.19.9.6076
Kuk MU, Kim JW, Lee YS, Cho KA, Park JT, Park SC (2019) Alleviation of senescence via ATM inhibition in accelerated aging models. Mol Cells 42:210–217. https://doi.org/10.14348/molcells.2018.0352
Chow HM, Cheng A, Song X, Swerdel MR, Hart RP, Herrup K (2019) ATM is activated by ATP depletion and modulates mitochondrial function through NRF1. J Cell Biol 218:909–928. https://doi.org/10.1083/jcb.201806197
Acknowledgements
We are thankful to Dr David Chen, UT Southwestern Medical Center, Dallas, TX, for providing us PcDNA-YFP-FLAG-WT and S1981A plasmids. FACS analysis was performed in the MD Anderson Cancer Center South Campus Flow Cytometry & Cell Sorting Core (supported by NCI Grant P30CA016672). We thank Stephanie Deming of Scientific Publications, Research Medical Library, MD Anderson Cancer Center, for editing the manuscript.
Funding
This work was supported in part by a grant from the CLL Global Research Foundation (to V.G.) and by the NIH/NCI through MD Anderson’s Cancer Center Support Grant, CA016672.
Author information
Authors and Affiliations
Contributions
AS conceptualized the study, designed experiments, generated and analyzed data, and wrote the manuscript. VG supervised the research project, analyzed data, obtained funding, and reviewed and edited the manuscript.
Corresponding authors
Ethics declarations
Conflict of interest
The authors declare no conflict of interest.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Electronic supplementary material
Below is the link to the electronic supplementary material.
Rights and permissions
About this article
Cite this article
Sarkar, A., Gandhi, V. Activation of ATM kinase by ROS generated during ionophore-induced mitophagy in human T and B cell malignancies. Mol Cell Biochem 476, 417–423 (2021). https://doi.org/10.1007/s11010-020-03917-1
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11010-020-03917-1