Deconstruction of the beaten Path-Sidestep interaction network provides insights into neuromuscular system development

Creators: Li, Hanqing; Watson, Ash; Olechwier, Agnieszka; Anaya, Michael; Sorooshyari, Siamak K.; Harnett, Dermott P.; Lee, Hyung-Kook (Peter); Vielmetter, Jost; Fares, Mario A.; Garcia, K. Christopher; Özkan, Engin; Labrador, Juan-Pablo; Zinn, Kai

Abstract

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.

Additional Information

© 2017 Li et al. This article is distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use and redistribution provided that the original author and source are credited. Received: 27 April 2017; Accepted: 28 July 2017; Published: 15 August 2017. This work was supported by NIH grants to KZ (R37 NS28182), and to EÖ (RO1 NS097161), by the Klingenstein-Simons Fellowship in the Neurosciences to EÖ, and by SFI grants 07/IN.1/B913 and 08/RFP/NSC1617 to J-P L. We thank Aref Arzan Zarin for preliminary genetic analysis. We thank Elena Armand and Suzanne Fisher for technical assistance, Maria Prats for preparation of AP supernatants for the experiments of Figs. S2 and S3, and Violana Nesterova for figure preparation. We thank Kaushiki Menon and Namrata Bali for help with larval staining. We acknowledge Dr. Elena Solomaha and the University of Chicago BioPhysics Core Facilities for training with and access to a Biacore 3000. We thank Laura Quintana Rio, Lalanti Venkatasubramanian, and Richard Mann (Columbia) for the Side-VI-T2A-GAL4 line. The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. Author Contributions: Hanqing Li, Conceptualization, Resources, Data curation, Formal analysis, Validation, Investigation, Writing—original draft, Writing—review and editing; Ash Watson, Hyung-Kook (Peter) Lee, Investi- gation, Visualization; Agnieszka Olechwier, Michael Anaya, Validation, Investigation, Methodology; Siamak K Sorooshyari, Software, Investigation, Methodology; Dermott P Harnett, Investigation, Methodology; Jost Vielmetter, Resources, Data curation, Funding acquisition, Validation, Investiga- tion, Methodology, Project administration; Mario A Fares, Conceptualization, Software, Investiga- tion, Methodology; K Christopher Garcia, Conceptualization, Resources, Project administration; Engin O¨ zkan, Conceptualization, Data curation, Formal analysis, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Project administration, Writing—review and editing; Juan-Pablo Labrador, Conceptualization, Resources, Data curation, Formal analysis, Supervision, Funding acquisition, Validation, Investigation, Visualization, Methodology, Project administration, Writing—review and editing; Kai Zinn, Conceptualization, Data curation, Formal analysis, Supervision, Funding acquisition, Investigation, Visualization, Methodology, Writing—original draft, Project administration, Writing—review and editing

Attached Files

Published - elife-28111-v3.pdf

Files

elife-28111-v3.pdf

Files (5.7 MB)

Name	Size	Download all
elife-28111-v3.pdf md5:0b5e6cca3e6b55478d1d190f57875180	5.7 MB	Preview Download

Additional details

	All versions	This version
Views	0	0
Downloads	0	0
Data volume	0 Bytes	0 Bytes