Targeted Expression of the Class II Phosphoinositide 3-Kinase in Drosophila melanogaster Reveals Lipid Kinase-Dependent Effects on Patterning and Interactions with Receptor Signaling Pathways

Abstract

Phosphoinositide 3-kinases (PI3Ks) can be divided into three distinct classes (I, II, and III) on the basis of their domain structures and the lipid signals that they generate. Functions have been assigned to the class I and class III enzymes but have not been established for the class II PI3Ks. We have obtained the first evidence for a biological function for a class II PI3K by expressing this enzyme during Drosophila melanogaster development and by using deficiencies that remove the endogenous gene. Wild-type and catalytically inactive PI3K_68D transgenes have opposite effects on the number of sensory bristles and on wing venation phenotypes induced by modified epidermal growth factor (EGF) receptor signaling. These results indicate that the endogenous PI3K_68D may act antagonistically to the EGF receptor-stimulated Ras-mitogen-activated protein kinase pathway and downstream of, or parallel to, the Notch receptor. A class II polyproline motif in PI3K_68D can bind the Drk adaptor protein in vitro, primarily via the N-terminal SH3 domain of Drk. Drk may thus be important for the localization of PI3K_68D, allowing it to modify signaling pathways downstream of cell surface receptors. The phenotypes obtained are markedly distinct from those generated by expression of the Drosophila class I PI3K, which affects growth but not pattern formation.

This work was supported by grants from the BBSRC and from the Royal Society and by the Ludwig Institute. E.H. is supported by a grant from the Swiss National Science Foundation.

We thank Kathy Matthews and the Bloomington Stock Center, Mandy Simcox, and Christian Lehner for the fly stocks used in this study; the Developmental Studies Hybridoma Bank for the 9E10 supernatant; Tony Pawson for the GST-Drk, GST-Drk^W36A, and GST-Drk^W189A constructs; Simon Woodcock and David Hughes for the SH3 domain of p120 RasGAP; Brenda Catelani, Alan Entwhistle, Krishna Pitrola, and Heather Phillips for technical assistance; Juan Riesgo-Escovar and Christoph Hugentobler for help in generating transgenic lines; Jenny Higgs for advice on imaging thoraxes; Carmen Coelho, María Domínguez, Enrique Martín-Blanco, and members of the Hafen lab, past and present, for useful discussions; and David Hughes for constructive criticism of the manuscript.