TY - JOUR
T1 - Coincident signals from GPCRs and receptor tyrosine kinases are uniquely transduced by PI3Kβ in myeloid cells
AU - Houslay, Daniel M.
AU - Anderson, Karen E.
AU - Chessa, Tamara
AU - Kulkarni, Suhasini
AU - Fritsch, Ralph
AU - Downward, Julian
AU - Backer, Jonathan M.
AU - Stephens, Len R.
AU - Hawkins, Phillip T.
N1 - Funding Information:
We would like to thank members of the Small Animal Breeding Unit at the Babraham Institute for animal husbandry; R. Williams (Medical Research Council Laboratory of Molecular Biology, Cambridge) for communicating essential information about the direct interaction between Gβγ and p110β; J. Clark for help with the MS analysis of lipids; A. Segonds-Pichon for statistical analysis; S. Walker and H. Okkenhaug (Babraham Institute Imaging Facility) for assistance with image analysis; and S. Suire and D. Gyori for helpful discussions. Funding: This work was supported by funding from the Biotechnology and Biological Sciences Research Council (BBSRC) UK (BB/J004456/1 to K.E.A., L.R.S., and P.T.H.), the Welcome Trust (WT085889MA to T.C. and S.K.), the NIH (GM112524 to J.M.B.), and the European Research Council (RASTARGET to J.D.). D.M.H. is a recipient of a BBSRC PhD studentship. Author contributions: K.E.A., D.M.H., S.K., and T.C. performed experiments; K.E.A., D.H., P.T.H., and L.R.S. analyzed data; R.F., J.D., and J.M.B. supplied important resources and information; P.T.H., K.E.A., and L.S. designed research; and P.T.H., K.E.A., T.C., and L.R.S. wrote the article. Competing interests: The authors declare that they have no competing interests. Data and materials availability: Use of the p110β KO and p110δ D910A-KI mice requires a material transfer agreement from the Ludwig Institute for Cancer Research.
Funding Information:
We would like to thank members of the Small Animal Breeding Unit at the Babraham Institute for animal husbandry; R. Williams (Medical Research Council Laboratory of Molecular Biology, Cambridge) for communicating essential information about the direct interaction between G?? and p110?; J. Clark for help with the MS analysis of lipids; A. Segonds-Pichon for statistical analysis; S. Walker and H. Okkenhaug (Babraham Institute Imaging Facility) for assistance with image analysis; and S. Suire and D. Gyori for helpful discussions. Funding: This work was supported by funding from the Biotechnology and Biological Sciences Research Council (BBSRC) UK (BB/J004456/1 to K.E.A., L.R.S., and P.T.H.), the Welcome Trust (WT085889MA to T.C. and S.K.), the NIH (GM112524 to J.M.B.), and the European Research Council (RASTARGET to J.D.). D.M.H. is a recipient of a BBSRC PhD studentship. Author contributions: K.E.A., D.M.H., S.K., and T.C. performed experiments; K.E.A., D.H., P.T.H., and L.R.S. analyzed data; R.F., J.D., and J.M.B. supplied important resources and information; P.T.H., K.E.A., and L.S. designed research; and P.T.H., K.E.A., T.C., and L.R.S. wrote the article. Competing interests: The authors declare that they have no competing interests. Data and materials availability: Use of the p110? KO and p110? D910A-KI mice requires a material transfer agreement from the Ludwig Institute for Cancer Research.
PY - 2016/8/16
Y1 - 2016/8/16
N2 - Class I phosphoinositide 3-kinases (PI3Ks) catalyze production of the lipid messenger phosphatidylinositol 3,4,5-trisphosphate (PIP3), which plays a central role in a complex signaling network regulating cell growth, survival, and movement. This network is overactivated in cancer and inflammation, and there is interest in determining the PI3K catalytic subunit (p110α, p110β, p110γ, or p110δ) that should be targeted in different therapeutic contexts. Previous studies have defined unique regulatory inputs for p110β, including direct interaction with Gβγ subunits, Rac, and Rab5. We generated mice with knock-in mutations of p110β that selectively blocked the interaction with Gβgγ and investigated its contribution to the PI3K isoform dependency of receptor tyrosine kinase (RTK) and G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptor (GPCR) responses in primary macrophages and neutrophils. We discovered a unique role for p110β in supporting synergistic PIP3 formation in response to the coactivation of macrophages by macrophage colony-stimulating factor (M-CSF) and the complement protein C5a. In contrast, we found partially redundant roles for p110α, p110β, and p110δ downstream of M-CSF alone and a nonredundant role for p110γ downstream of C5a alone. This role for p110β completely depended on direct interaction with Gβγ, suggesting that p110β transduces GPCR signals in the context of coincident activation by an RTK. The p110β-Gβγ interaction was also required for neutrophils to generate reactive oxygen species in response to the Fcγ receptor-dependent recognition of immune complexes and for their β2 integrin-mediated adhesion to fibrinogen or poly-RGD+, directly implicating heterotrimeric G proteins in these two responses.
AB - Class I phosphoinositide 3-kinases (PI3Ks) catalyze production of the lipid messenger phosphatidylinositol 3,4,5-trisphosphate (PIP3), which plays a central role in a complex signaling network regulating cell growth, survival, and movement. This network is overactivated in cancer and inflammation, and there is interest in determining the PI3K catalytic subunit (p110α, p110β, p110γ, or p110δ) that should be targeted in different therapeutic contexts. Previous studies have defined unique regulatory inputs for p110β, including direct interaction with Gβγ subunits, Rac, and Rab5. We generated mice with knock-in mutations of p110β that selectively blocked the interaction with Gβgγ and investigated its contribution to the PI3K isoform dependency of receptor tyrosine kinase (RTK) and G protein (heterotrimeric guanine nucleotide-binding protein)-coupled receptor (GPCR) responses in primary macrophages and neutrophils. We discovered a unique role for p110β in supporting synergistic PIP3 formation in response to the coactivation of macrophages by macrophage colony-stimulating factor (M-CSF) and the complement protein C5a. In contrast, we found partially redundant roles for p110α, p110β, and p110δ downstream of M-CSF alone and a nonredundant role for p110γ downstream of C5a alone. This role for p110β completely depended on direct interaction with Gβγ, suggesting that p110β transduces GPCR signals in the context of coincident activation by an RTK. The p110β-Gβγ interaction was also required for neutrophils to generate reactive oxygen species in response to the Fcγ receptor-dependent recognition of immune complexes and for their β2 integrin-mediated adhesion to fibrinogen or poly-RGD+, directly implicating heterotrimeric G proteins in these two responses.
UR - http://www.scopus.com/inward/record.url?scp=84983468958&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84983468958&partnerID=8YFLogxK
U2 - 10.1126/scisignal.aae0453
DO - 10.1126/scisignal.aae0453
M3 - Article
C2 - 27531651
AN - SCOPUS:84983468958
SN - 1945-0877
VL - 9
JO - Science Signaling
JF - Science Signaling
IS - 441
M1 - ra82
ER -