Identification of protein phosphorylation targets of S1b/Wnt11 signalling during zebrafish gastrulation

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We have shown that Wnt11/Slb function is required for normal cell movement during zebrafish gastrulation. In this process, Wnt11/Slb is thought to signal through a pathway similar to the one used for establishment of planar cell polarity in Drosophila. Key components of this signalling pathway in Drosophila are the small GTPases rho, rac and cdc42 that can function through direct phosphorylation of effector proteins. To identify targets of Wnt11/Slb signalling during zebrafish gastrulation, we have therefore decided to screen for proteins being phosphorylated in the presence of Wnt11/Slb signals. We will identify such proteins using two-dimensional gel electrophoresis and mass-spectrometry (in collaboration with the mass-spectrometry laboratory of Dr Shevchenko in our institute). The function of these proteins in zebrafish gastrulation will subsequently be tested using gain- and loss-of-function techniques. The identification of target proteins of Wnt11/Slb signalling will lead to a better understanding of the molecular mechanisms that control cell movement during zebrafish gastrulation.