DNA-PKcs Controls an Endosomal Signaling Pathway for a Proinflammatory Response by Natural Killer Cells
Under the Surface
In addition to serving as compartments in which signaling by internalized receptors is terminated, endosomes provide venues in which other receptors continue to signal or even trigger alternative pathways. Indeed, some receptors that fail to signal at the membrane function efficiently in endosomes. CD158d, an endosome-resident, immunoglobulin-like receptor found mostly in natural killer (NK) cells, triggers a proinflammatory and proangiogenic pathway in response to soluble human leukocyte antigen G (HLA-G). Uptake of fetal trophoblast–derived HLA-G by maternal NK cells may trigger the vascular remodeling required to establish an effective blood supply to the fetus, but how CD158d stimulates this response is unclear. Rajagopalan et al. found that endosomal CD158d activated nuclear factor κB (NF-κB) through a mechanism that depended on the kinase Akt, but not phosphatidylinositol 3-kinase (PI3K). In addition, CD158d associated with the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), which phosphorylated Akt at the endosome. In addition to expanding the known roles of DNA-PKcs, this study also highlights a PI3K-independent, endosomal function for Akt that is required for a proinflammatory response in NK cells.
Abstract
Endosomes are emerging as specialized signaling compartments that endow receptors with distinct signaling properties. The diversity of endosomal signaling pathways and their contribution to various biological responses is still unclear. CD158d, which is also known as the killer cell immunoglobulin-like receptor (KIR) 2DL4 (KIR2DL4), is an endosome-resident receptor in natural killer (NK) cells that stimulates the release of a unique set of proinflammatory and proangiogenic mediators in response to soluble human leukocyte antigen G (HLA-G). Here, we identified the CD158d signaling cascade. In response to soluble agonist antibody or soluble HLA-G, signaling by CD158d was dependent on the activation of nuclear factor κB (NF-κB) and the serine-threonine kinase Akt. CD158d associated with the catalytic subunit of DNA-dependent protein kinase (DNA-PKcs), promoted the recruitment of Akt to endosomes, and stimulated the DNA-PKcs–dependent phosphorylation of Akt. The sequential requirement for DNA-PKcs, Akt, and NF-κB in signaling by CD158d delineates a previously uncharacterized endosomal signaling pathway for a proinflammatory response in NK cells.
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Supplementary Material
Summary
Fig. S1. The CD158d-gp49 chimera traffics to the same endosomal compartments as does wild-type CD158d.
Fig. S2. Stimulation of CD158d in resting NK cells results in the phosphorylation of Akt at Ser473, but not Thr308.
Fig. S3. Inhibition of the CD158d-dependent secretion of IFN-γ and IL-8.
Fig. S4. CD158d colocalizes with Rab5 and Rab5Q79L in endosomes.
Fig. S5. Akt is present in endosomes that contain CD158d and Rab5Q79L.
Fig. S6. Colocalization profiles of CD158d, Rab5Q79L, and Akt.
Table S1. Compounds tested for their ability to inhibit CD158d-dependent secretion of IFN-γ by resting NK cells.
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Information & Authors
Information
Published In
Science Signaling
Volume 3 | Issue 110
February 2010
February 2010
Copyright
Copyright © 2010, American Association for the Advancement of Science.
Submission history
Received: 15 June 2009
Accepted: 5 February 2010
Acknowledgments
Acknowledgments: We thank W. Lane (Harvard Microchemistry) for mass spectrometry analysis and H. Young, U. Siebenlist, J. Bonifacino, Y. Bryceson, B. P. Chen, and D. J. Chen for plasmids.
Funding: This work was supported by the Intramural Research Program of the NIH, National Institute of Allergy and Infectious Diseases.
Author contributions: S.R. and E.O.L. designed the experiments. S.R. and M.W.M. performed experiments and interpreted results. I.J. prepared and characterized a recombinant protein. S.R. and E.O.L. analyzed data and wrote the paper.
Competing interests: S.R. and E.O.L. have U.S. Patent 7,435,801: “Antibodies and other ligands directed against KIR2DL4 receptor for production of interferon gamma.”
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