REL 2021-02-02 2021-04-06 2022-02-07 0000-0002-4847-5529 Tomas Malinauskas Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford Roosevelt Drive Oxford Oxfordshire United Kingdom OX3 7BN 0000-0002-6635-3621 Christian Siebold Division of Structural Biology, Wellcome Centre for Human Genetics, University of Oxford Roosevelt Drive Oxford Oxfordshire United Kingdom OX3 7BN Robinson RA Griffiths SC van de Haar LL Malinauskas T van Battum EY Zelina P Schwab RA Karia D Malinauskaite L Brignani S van den Munkhof M Dudukcu O De Ruiter AA Van den Heuvel DMA Bishop B Elegheert J Aricescu AR Pasterkamp RJ Siebold C 1.2 10.6019/EMPIAR-10637 EMDB EMD-12286 Robinson RA Griffiths SC van de Haar LL Malinauskas T van Battum EY Zelina P Schwab RA Karia D Malinauskaite L Brignani S van den Munkhof M Dudukcu O De Ruiter AA Van den Heuvel DMA Bishop B Elegheert J Aricescu AR Pasterkamp RJ Siebold C Cell Cell 2021 English 10.1016/j.cell.2021.02.045 33740419

During cell migration or differentiation, cell surface receptors are exposed to different ligands. However, how the signals from simultaneously presented extracellular ligands are integrated remains incompletely understood. Neogenin (NEO1) acts as an attractive guidance receptor upon Netrin-1 (NET1) binding but mediates repulsion via Repulsive Guidance Molecules (RGMs). Here, we identify a mechanism of signal integration through the formation of a ternary NEO1-NET1-RGM complex triggering reciprocal silencing of downstream signaling. We determine structures of the NEO1-NET1-RGM complex, revealing a defined ‘trimer-of-trimers’ super-assembly, and demonstrate that this complex can exist in the cell membrane. Formation of this super-assembly results in inhibition of RGMA-NEO1-mediated growth cone collapse and RGMA- or NET1-NEO1-mediated neuron migration, by preventing signalling-compatible RGM-NEO1 complexes and NET1-induced NEO1 ectodomain clustering. These results illustrate how simultaneous binding of ligands with opposing functions to a single receptor does not lead to competition for binding, but to formation of a super-complex that diminishes their functional outputs.

Unaligned multi-frame micrographs of the ternary Netrin 1-Neogenin 1-Repulsive Guidance Molecule B complex /data micrographs - multiframe TIFF TIFF 1635 40 SIGNED 16 BIT INTEGER 4096 0.85 4096 0.85