Deconstruction of the beaten Path-Sidestep interaction network provides insights into neuromuscular system development
Hanqing Li,
Ash Watson,
Agnieszka Olechwier,
Michael Anaya,
Siamak K Sorooshyari,
Dermott P Harnett,
Hyung-Kook (Peter) Lee,
Jost Vielmetter,
Mario A Fares,
K Christopher Garcia,
Engin Özkan,
Juan-Pablo Labrador,
Kai Zinn
Affiliations
Hanqing Li
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States
Ash Watson
Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland; Institute of Neuroscience, Trinity College Dublin, University of Dublin, Dublin, Ireland
Agnieszka Olechwier
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, United States
Michael Anaya
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States
Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland; Institute of Neuroscience, Trinity College Dublin, University of Dublin, Dublin, Ireland
Hyung-Kook (Peter) Lee
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States
Jost Vielmetter
Division of Biology and Biological Engineering, California Institute of Technology, Pasadena, United States
Mario A Fares
Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland; Department of Abiotic Stress, Group of Integrative and Systems Biology, Instituto de Biología Molecular y Celular de Plantas (CSIC-Universidad Politécnica de Valencia), Valencia, Spain
Department of Molecular and Cellular Physiology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, United States; Department of Structural Biology, Howard Hughes Medical Institute, Stanford University School of Medicine, Stanford, United States
Department of Biochemistry and Molecular Biology, University of Chicago, Chicago, United States
Juan-Pablo Labrador
Smurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland; Institute of Neuroscience, Trinity College Dublin, University of Dublin, Dublin, Ireland
An ‘interactome’ screen of all Drosophila cell-surface and secreted proteins containing immunoglobulin superfamily (IgSF) domains discovered a network formed by paralogs of Beaten Path (Beat) and Sidestep (Side), a ligand-receptor pair that is central to motor axon guidance. Here we describe a new method for interactome screening, the Bio-Plex Interactome Assay (BPIA), which allows identification of many interactions in a single sample. Using the BPIA, we ‘deorphanized’ four more members of the Beat-Side network. We confirmed interactions using surface plasmon resonance. The expression patterns of beat and side genes suggest that Beats are neuronal receptors for Sides expressed on peripheral tissues. side-VI is expressed in muscle fibers targeted by the ISNb nerve, as well as at growth cone choice points and synaptic targets for the ISN and TN nerves. beat-V genes, encoding Side-VI receptors, are expressed in ISNb and ISN motor neurons.
