PHD3-dependent hydroxylation of HCLK2 promotes the DNA damage response
- 1 August 2012
- journal article
- Published by American Society for Clinical Investigation in JCI Insight
- Vol. 122 (8), 2827-2836
- https://doi.org/10.1172/JCI62374
Abstract
The DNA damage response (DDR) is a complex regulatory network that is critical for maintaining genome integrity. Posttranslational modifications are widely used to ensure strict spatiotemporal control of signal flow, but how the DDR responds to environmental cues, such as changes in ambient oxygen tension, remains poorly understood. We found that an essential component of the ATR/CHK1 signaling pathway, the human homolog of the Caenorhabditis elegans biological clock protein CLK-2 (HCLK2), associated with and was hydroxylated by prolyl hydroxylase domain protein 3 (PHD3). HCLK2 hydroxylation was necessary for its interaction with ATR and the subsequent activation of ATR/CHK1/p53. Inhibiting PHD3, either with the pan-hydroxylase inhibitor dimethyloxaloylglycine (DMOG) or through hypoxia, prevented activation of the ATR/CHK1/p53 pathway and decreased apoptosis induced by DNA damage. Consistent with these observations, we found that mice lacking PHD3 were resistant to the effects of ionizing radiation and had decreased thymic apoptosis, a biomarker of genomic integrity. Our identification of HCLK2 as a substrate of PHD3 reveals the mechanism through which hypoxia inhibits the DDR, suggesting hydroxylation of HCLK2 is a potential therapeutic target for regulating the ATR/CHK1/p53 pathway.Keywords
This publication has 33 references indexed in Scilit:
- Prolyl Hydroxylase PHD3 Enhances the Hypoxic Survival and G1 to S Transition of Carcinoma CellsPLOS ONE, 2011
- Chk2-deficient mice exhibit radioresistance and defective p53-mediated transcriptionThe EMBO Journal, 2002
- Sequence Determinants in Hypoxia-inducible Factor-1α for Hydroxylation by the Prolyl Hydroxylases PHD1, PHD2, and PHD3Published by Elsevier BV ,2002
- C. elegans EGL-9 and Mammalian Homologs Define a Family of Dioxygenases that Regulate HIF by Prolyl HydroxylationCell, 2001
- HIFα Targeted for VHL-Mediated Destruction by Proline Hydroxylation: Implications for O 2 SensingScience, 2001
- Targeting of HIF-α to the von Hippel-Lindau Ubiquitylation Complex by O 2 -Regulated Prolyl HydroxylationScience, 2001
- The DNA-repair protein AlkB, EGL-9, and leprecan define new families of 2-oxoglutarate- and iron-dependent dioxygenasesGenome Biology, 2001
- SM-20 Is a Novel Mitochondrial Protein That Causes Caspase-dependent Cell Death in Nerve Growth Factor-dependent NeuronsPublished by Elsevier BV ,2001
- p53AIP1, a Potential Mediator of p53-Dependent Apoptosis, and Its Regulation by Ser-46-Phosphorylated p53Cell, 2000
- HIF-1: mediator of physiological and pathophysiological responses to hypoxiaJournal of Applied Physiology, 2000