The DNA damage pathway regulates innate immune system ligands of the NKG2D receptor

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Some stimulatory receptors of the innate immune system, such as the NKG2D receptor (also called KLRK1) expressed by natural killer cells and activated CD8+T cells, recognize self-molecules that are upregulated in diseased cells by poorly understood mechanisms1. Here we show that mouse and human NKG2D ligands are upregulated in non-tumour cell lines by genotoxic stress and stalled DNA replication, conditions known to activate a major DNA damage checkpoint pathway initiated by ATM (ataxia telangiectasia, mutated) or ATR (ATM- and Rad3-related) protein kinases2. Ligand upregulation was prevented by pharmacological or genetic inhibition of ATR, ATM or Chk1 (a downstream transducer kinase in the pathway). Furthermore, constitutive ligand expression by a tumour cell line was inhibited by targeting short interfering RNA to ATM, suggesting that ligand expression in established tumour cells, which often harbour genomic irregularities, may be due to chronic activation of the DNA damage response pathway. Thus, the DNA damage response, previously shown to arrest the cell cycle and enhance DNA repair functions, or to trigger apoptosis, may also participate in alerting the immune system to the presence of potentially dangerous cells.

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Figure 1: NKG2D ligand upregulation is associated with tumorigenesis, not with transformation per se.
Figure 2: NKG2D ligands are induced by DNA-damaging agents and DNA synthesis inhibitors.
Figure 3: Kinetics of the upregulation of NKG2D ligands and phosphorylation of the Chk1 kinase.
Figure 4: Ligand upregulation in aphidicolin-treated fibroblasts is dependent on ATR function.
Figure 5: Role of Chk1 in expression of NKG2D ligands in aphidicolin-treated fibroblasts and tumour cell lines.


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We thank J. Thorner and S. Linn for discussions; Y. Natanzon and L. Zhang for technical assistance; H. Nolla for assistance with flow cytometry; and L. Fritz-Laylin for help with transductions. This work was supported by a National Institutes of Health grant to D.H.R. and an award from the Prostate Cancer Foundation.

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Correspondence to David H. Raulet.

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Supplementary information

Supplementary Figure S1

Specific induction of NKG2D ligands by treatments that activate the DNA damage response. (PDF 692 kb)

Supplementary Figure S2

Real time RT-PCR quantitation of Rae1 and MULT1 transcript levels in cell lines treated with aphidicolin. (PDF 483 kb)

Supplementary Figure S3

Cell cycle status of ATR-deleted fibroblasts. (PPT 141 kb)

Supplementary Figure S4

ATR/ATM inhibitors prevent NKG2D ligand upregulation. (PPT 319 kb)

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Gasser, S., Orsulic, S., Brown, E. et al. The DNA damage pathway regulates innate immune system ligands of the NKG2D receptor. Nature 436, 1186–1190 (2005) doi:10.1038/nature03884

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