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Science 287 (5455): 1037-1040

Copyright © 2000 by the American Association for the Advancement of Science

Polarization of Chemoattractant Receptor Signaling During Neutrophil Chemotaxis

Guy Servant, 1* Orion D. Weiner, 12* Paul Herzmark, 1 Tamás Balla, 3 John W. Sedat, 2 Henry R. Bourne 1dagger

Morphologic polarity is necessary for chemotaxis of mammalian cells. As a probe of intracellular signals responsible for this asymmetry, the pleckstrin homology domain of the AKT protein kinase (or protein kinase B), tagged with the green fluorescent protein (PHAKT-GFP), was expressed in neutrophils. Upon exposure of cells to chemoattractant, PHAKT-GFP is recruited selectively to membrane at the cell's leading edge, indicating an internal signaling gradient that is much steeper than that of the chemoattractant. Translocation of PHAKT-GFP is inhibited by toxin-B from Clostridium difficile, indicating that it requires activity of one or more Rho guanosine triphosphatases.

1 Department of Cellular and Molecular Pharmacology and
2 Department of Biochemistry and Biophysics, University of California San Francisco, San Francisco, CA 94143, USA.
3 Endocrinology and Reproduction Research Branch, National Institutes of Child Health and Human Development, National Institutes of Health, Bethesda, MD 20892-4510, USA.
*   These authors contributed equally to this work.

dagger    To whom correspondence should be addressed. E-mail: bourne{at}cmp.ucsf.edu

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Science 312, 443-446
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Single-molecule analysis of chemoattractant-stimulated membrane recruitment of a PH-domain-containing protein.
S. Matsuoka, M. Iijima, T. M. Watanabe, H. Kuwayama, T. Yanagida, P. N. Devreotes, and M. Ueda (2006)
J. Cell Sci. 119, 1071-1079
   Abstract »    Full Text »    PDF »
Neutrophil polarization: Spatiotemporal dynamics of RhoA activity support a self-organizing mechanism..
K. Wong, O. Pertz, K. Hahn, and H. Bourne (2006)
PNAS 103, 3639-3644
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Neutrophil chemorepulsion in defined interleukin-8 gradients in vitro and in vivo.
W. G. Tharp, R. Yadav, D. Irimia, A. Upadhyaya, A. Samadani, O. Hurtado, S-Y. Liu, S. Munisamy, D. M. Brainard, M. J. Mahon, et al. (2006)
J. Leukoc. Biol. 79, 539-554
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Rho GEF Lsc is required for normal polarization, migration, and adhesion of formyl-peptide-stimulated neutrophils.
S. A. Francis, X. Shen, J. B. Young, P. Kaul, and D. J. Lerner (2006)
Blood 107, 1627-1635
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Localised PtdIns(3,4,5)P3 or PtdIns(3,4)P2 at the phagocytic cup is required for both phagosome closure and Ca2+ signalling in HL60 neutrophils.
S. Dewitt, W. Tian, and M. B. Hallett (2006)
J. Cell Sci. 119, 443-451
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The Shwachman-Bodian-Diamond syndrome gene encodes an RNA-binding protein that localizes to the pseudopod of Dictyostelium amoebae during chemotaxis.
D. Wessels, T. Srikantha, S. Yi, S. Kuhl, L. Aravind, and D. R. Soll (2006)
J. Cell Sci. 119, 370-379
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