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Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion

Nature Immunology volume 7, pages 919–928 (2006)Cite this article

Abstract

RAPL, a protein that binds the small GTPase Rap1, is required for efficient immune cell trafficking. Here we have identified the kinase Mst1 as a critical effector of RAPL. RAPL regulated the localization and kinase activity of Mst1. 'Knockdown' of the gene encoding Mst1 demonstrated its requirement for the induction of both a polarized morphology and integrin LFA-1 clustering and adhesion triggered by chemokines and T cell receptor ligation. RAPL and Mst1 localized to vesicular compartments and dynamically translocated with LFA-1 to the leading edge upon Rap1 activation, suggesting a regulatory function for the RAPL-Mst1 complex in intracellular transport of LFA-1. Our study demonstrates a previously unknown function for Mst1 of relaying the Rap1-RAPL signal to induce cell polarity and adhesion of lymphocytes.

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Figure 1: Association of RAPL and MST1.
Figure 2: Mst1 activation by RAPL and Rap1.
Figure 3: MST1 induces cell adhesion to ICAM-1.
Figure 4: Mst1 'knockdown' abrogates cell polarization and adhesion.
Figure 5: Mst1 'knockdown' abrogates TCR-mediated adhesion and immunological synapse formation.
Figure 6: Mst1 'knockdown' in mouse T lymphocytes abrogates the cell adhesion triggered by CCL21 stimulation and TCR ligation.
Figure 7: Subcellular distribution of Mst1 and RAPL.
Figure 8: Rapid intracellular translocation of MST1, RAPL and LFA-1.

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Acknowledgements

We thank N. Shimomura, C. Tanaka and R. Hamaguchi for technical assistance. Supported by a grant-in-aid from the Ministry of Education, Science, Sports, and Culture of Japan; Uehara Memorial Foundation; and Toray Science Foundation.

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Authors and Affiliations

  1. Department of Molecular Genetics, Institute of Biomedical Science, Kansai Medical University, Osaka, 570-8506, Japan

    Koko Katagiri & Tatsuo Kinashi

  2. Center Research Institute, Ishihara Sangyo Kaisha, Kusatsu, 525-0025, Shiga, Japan

    Masashi Imamura

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Contributions

K.K. and T.K. contributed to discussions of experimental design, data analysis and manuscript preparation and did all experimental studies unless otherwise indicated; M.I. did immunoelectron microscopy.

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Correspondence to Tatsuo Kinashi.

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Supplementary information

Supplementary Fig. 1

Localization with Mst1 in T cells. (PDF 452 kb)

Supplementary Fig. 2

Pseudo-color image of LFA-1 intensities of Z-stack confocal images of the cell (asterisk) indicated in Fig. 3(d). (PDF 353 kb)

Supplementary Fig. 3

Affinity regulation by Mst1. (PDF 692 kb)

Supplementary Fig. 4

Co-localization analysis. (PDF 337 kb)

Supplementary Fig. 5

Localization of Mst1 in immunological synapse. (PDF 372 kb)

Supplementary Fig. 6

Immunoelectron microscopy of RAPL in 3A9 T cells. (PDF 1735 kb)

Supplementary Fig. 7

Relocation of MST-mRFP toward the leading edge. (PDF 471 kb)

Supplementary Fig. 8

Immunostaining for tubulin. (PDF 377 kb)

Supplementary Fig. 9

Distribution of β2-mRFP. (PDF 423 kb)

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Katagiri, K., Imamura, M. & Kinashi, T. Spatiotemporal regulation of the kinase Mst1 by binding protein RAPL is critical for lymphocyte polarity and adhesion. Nat Immunol 7, 919–928 (2006). https://doi.org/10.1038/ni1374

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