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The lymphocyte function-associated antigen-1 receptor costimulates plasma membrane Ras via phospholipase D2

Nature Cell Biology volume 9, pages 713–719 (2007)Cite this article

Abstract

Ras activation as a consequence of antigen receptor (T-cell receptor; TCR) engagement on T lymphocytes is required for T-cell development, selection and function. Lymphocyte function-associated antigen-1 (LFA-1) mediates lymphocyte adhesion, stabilization of the immune synapse and bidirectional signalling. Using a fluorescent biosensor we found that TCR activation with or without costimulation of CD28 led to activation of Ras only on the Golgi apparatus, whereas costimulation with LFA-1 induced Ras activation on both the Golgi and the plasma membrane. Ras activation on both compartments required RasGRP1, an exchange factor regulated by calcium and diacylglycerol (DAG), but phospholipase C (PLC) activity was required only for activation on the Golgi. Engagement of LFA-1 increased DAG levels at the plasma membrane by stimulating phospholipase D (PLD). PLD2 and phosphatidic acid phosphatase (PAP) were required for Ras activation on the plasma membrane. Thus, LFA-1 acts through PLD2 to reshape the pattern of Ras activation downstream of the TCR.

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Figure 1: Costimulation of LFA-1 is required for TCR activation of Ras at the plasma membrane.
Figure 2: LFA-1 costimulates TCR activation of the Ras/MAPK pathway.
Figure 3: Costimulation through LFA-1 is required to elevate DAG levels and recruit RasGRP1 to the plasma membrane.
Figure 4: Costimulation of plasma membrane Ras activation by LFA-1 does not require PLCγ1.
Figure 5: Costimulation of plasma membrane Ras activation by LFA-1 requires PLD2.

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Acknowledgements

We thank J. Stone for generously supplying Ras–GRP1-deficient mice. We thank M. Frohman for PLD expression plasmids. We thank Y. Nozawa for antibodies against PLD2. This work was supported by grants from the National Institutes of Health and the Arthritis National Research Foundation.

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Authors and Affiliations

  1. Department of Medicine, NYU School of Medicine, New York, 10016, NY, USA

    Adam Mor & Mark R. Philips

  2. Department of Pathology, NYU School of Medicine, New York, 10016, NY, USA

    Gabriele Campi & Michael L. Dustin

  3. The Skirball Institute of Biomolecular Medicine, NYU School of Medicine, New York, 10016, NY, USA

    Gabriele Campi & Michael L. Dustin

  4. Department of Pharmacology, SUNY Stony Brook, 11794, NY, USA

    Guangwei Du

  5. Department of Biological Sciences, Hunter College, City University of New York, New York, 10021, NY, USA

    Yang Zheng & David A. Foster

  6. Department of Cell Biology, NYU School of Medicine, New York, 10016, NY, USA

    Mark R. Philips

  7. Department of Pharmacology NYU School of Medicine, New York, 10016, NY, USA

    Mark R. Philips

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  1. Adam Mor
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Contributions

A.M. and M.R.P. designed and analysed all experiments and prepared the manuscript. A.M. performed all experimental work. G.C. and M.L.D. assisted with ICAM-1 stimulation and T-cell imaging. G.D. assisted with PLD2 knockdown. Y.Z. and D.A.F. assisted with the PLD assay. M.L.D. and D.A.F. contributed to the discussion.

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Correspondence to Mark R. Philips.

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Mor, A., Campi, G., Du, G. et al. The lymphocyte function-associated antigen-1 receptor costimulates plasma membrane Ras via phospholipase D2. Nat Cell Biol 9, 713–719 (2007). https://doi.org/10.1038/ncb1592

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