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Gβγs and the Ras binding domain of p110γ are both important regulators of PI3Kγ signalling in neutrophils

Nature Cell Biology volume 8, pages 1303–1309 (2006)Cite this article

Abstract

Through their ability to regulate production of the key lipid messenger PtdIns(3,4,5)P3, the class I phosphatidylinositol-3-OH kinases (PI(3)Ks) support many critical cell responses1,2. They, in turn, can be regulated by cell-surface receptors through signals acting on either their adaptor subunits (for example, through phosphotyrosine or Gβγs) or their catalytic subunits (for example, through GTP-Ras). The relative significance of these controlling inputs is undefined in vivo. Here, we have studied the roles of Gβγs, the adaptor p101, Ras and the Ras binding domain (RBD) in the control of the class I PI(3)K, PI(3)Kγ, in mouse neutrophils. Loss of p101 leads to major reductions in the accumulation of PtdIns(3,4,5)P3, activation of protein kinase B (PKB) and in migration towards G-protein activating ligands in vitro, and to an aseptically inflamed peritoneum in vivo. Loss of sensitivity of PI(3)Kγ to Ras unexpectedly caused similar reductions, but additionally caused a substantial loss in production of reactive oxygen species (ROS). We conclude that Gβγs, p101 and the Ras–RBD interaction all have important roles in the regulation of PI(3)Kγ in vivo and that they can simultaneously, but differentially, control distinct PI(3)Kγ effectors.

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Figure 1: Expression and distributions of PI(3)K subunits in neutrophils isolated from p101−/− and p110γDASAA/DASAA mice.
Figure 2: Accumulation of PtdIns(3,4,5)P3 is reduced in neutrophils isolated from p110γDASAA/DASAA (p110γD/D) and p101−/− mice.
Figure 3: Phosphorylation of Ser 473-PKB in response to chemoattractants is reduced in neutrophils from p101−/− and p110γDASAA/DASAA mice.
Figure 4: The migration of neutrophils from mice lacking p110γ or p101 or expressing p110γDASAA/DASAA is substantially reduced.
Figure 5: Production of ROS in response to chemoattractants is reduced in neutrophils from p110γDASAA/DASAA but not p101−/− mice.

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Acknowledgements

PI(3)Kγ mice were provided by M. Wymann. Thanks to M. Skynner for help in the initial isolation of p101 genomic clones and thanks also to T. Green for use of a blood-cell counter. Thanks to P. Arnaud for design of PCR primers and A. Segonds-Pichon for help with statistics. C.E. was a Beit fellow. A.M.C. was supported by a Wellcome Trust Intermediate fellowship. Different parts of this work were funded by grants from the Biotechnology and Biological Sciences Research council (BBSRC) and Cancer Research UK (CRUK).

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Author notes

  1. Chris D. Ellson

    Present address: Centre for Cancer Research, 40 Ames Street, Cambridge, MA 02139, USA

  2. Sabine Suire and Alison M. Condliffe: These authors contributed equally to this work.

  3. Phillip T. Hawkins and Len Stephens: These authors contributed equally to this work.

Authors and Affiliations

  1. The Babraham Institute, Babraham, CB22 3AT, Cambs, UK

    Sabine Suire, Alison M. Condliffe, G. John Ferguson, Chris D. Ellson, Hervé Guillou, Keith Davidson, Heidi Welch, John Coadwell, Martin Turner, Phillip T. Hawkins & Len Stephens

  2. Respiratory Medicine Division, University of Cambridge School of Clinical Medicine, Addenbrooke's and Papworth Hospitals, Cambridge, UK

    Alison M. Condliffe & Edwin R. Chilvers

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Suire, S., Condliffe, A., Ferguson, G. et al. Gβγs and the Ras binding domain of p110γ are both important regulators of PI3Kγ signalling in neutrophils. Nat Cell Biol 8, 1303–1309 (2006). https://doi.org/10.1038/ncb1494

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