Nckβ Interacts with Tyrosine-Phosphorylated Disabled 1 and Redistributes in Reelin-Stimulated Neurons

Abstract

The tyrosine phosphorylation sites of the Disabled 1 (Dab1) docking protein are essential for the transmission of the Reelin signal, which regulates neuronal placement. Here we identify Nckβ as a phosphorylation-dependent, Dab1-interacting protein. The SH2 domain of Nckβ but not Nckα binds Dab1 phosphorylated on the Reelin-regulated site, Y220, or on Y232. Nckβ is coexpressed with Dab1 in the developing brain and in cultured neurons, where Reelin stimulation leads to the redistribution of Nckβ from the cell soma into neuronal processes. We found that tyrosine-phosphorylated Dab1 in synergy with Nckβ disrupts the actin cytoskeleton in transfected cells. In Drosophila melanogaster, exogenous expression of mouse Dab1 causes tyrosine phosphorylation site-dependent morphological changes in the compound eye. This phenotype is enhanced by overexpression of the Drosophila Nck protein Dock, suggesting a conserved interaction between the Disabled and Nck family members. We suggest a model in which Dab1 phosphorylation leads to the recruitment of Nckβ to the membrane, where it acts to remodel the actin cytoskeleton.

ACKNOWLEDGMENTS

We gratefully acknowledge Jon Cooper for support during the initial phase of this project and continuing encouragement, advice, and thoughtful comments on the manuscript. We appreciate the numerous gifts of reagents from Wei Li, Mark Henkemeyer, Chad Cowan, and Laurent Seroude. We value the many discussions on this work from the members of the Neurogenetics Branch. Technical support was provided by Jim Nagle with the NINDS sequencing facility.

This work was supported by NINDS intramural funds, an HHMI physician postdoctoral research fellowship to A.G., and HHMI-NIH research scholarships to S.M. and P.O.