Tetraspanin-3 is an organizer of the multi-subunit Nogo-A signaling complex.
To ensure precision and specificity of ligand-receptor-induced signaling, co-receptors and modulatory factors play important roles. The membrane-bound ligand Nogo-A (an isoform encoded by RTN4) induces inhibition of neurite outgrowth, cell spreading, adhesion and migration through multi-subunit receptor complexes. Here, we identified the four-transmembrane-spanning protein tetraspanin-3 (TSPAN3) as a new modulatory co-receptor for the Nogo-A inhibitory domain Nogo-A-Δ20. Single-molecule tracking showed that TSPAN3 molecules in the cell membrane reacted to binding of Nogo-A with elevated mobility, which was followed by association with the signal-transducing Nogo-A receptor sphingosine-1-phosphate receptor 2 (S1PR2). Subsequently, TSPAN3 was co-internalized as part of the Nogo-A-ligand-receptor complex into early endosomes, where it subsequently separated from Nogo-A and S1PR2 to be recycled to the cell surface. The functional importance of the Nogo-A-TSPAN3 interaction is shown by the fact that knockdown of TSPAN3 strongly reduced the Nogo-A-induced S1PR2 clustering, RhoA activation, cell spreading and neurite outgrowth inhibition. In addition to the modulatory functions of TSPAN3 on Nogo-A-S1PR2 signaling, these results illustrate the very dynamic spatiotemporal reorganizations of membrane proteins during ligand-induced receptor complex organization.
Brain Research Institute, University of Zurich and Dept. of Health Sciences & Technology, ETH Zurich, Zurich 8057, Switzerland thiede@hifo.uzh.ch schwab@hifo.uzh.ch.
Nina K
NK
Thiede-Stan
Brain Research Institute, University of Zurich and Dept. of Health Sciences & Technology, ETH Zurich, Zurich 8057, Switzerland.
Björn
B
Tews
Institute of Biochemistry and Laboratory of Physical Chemistry, ETH Zurich, Zurich 8093, Switzerland.
David
D
Albrecht
Brain Research Institute, University of Zurich and Dept. of Health Sciences & Technology, ETH Zurich, Zurich 8057, Switzerland.
Zorica
Z
Ristic
Institute of Biochemistry and Laboratory of Physical Chemistry, ETH Zurich, Zurich 8093, Switzerland.
Helge
H
Ewers
Brain Research Institute, University of Zurich and Dept. of Health Sciences & Technology, ETH Zurich, Zurich 8057, Switzerland thiede@hifo.uzh.ch schwab@hifo.uzh.ch.
Martin E
ME
Schwab
D000818:Animals; D002462:Cell Membrane / Q000378:metabolism; D011992:Endosomes / Q000378:metabolism; D047468:Immunoprecipitation; D051379:Mice; D009185:Myelin Proteins / Q000235:genetics / Q000378:metabolism*; D041681:NIH 3T3 Cells; D000070798:Nogo Proteins; D049349:Receptors, Lysosphingolipid / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology; D059470:Tetraspanins / Q000235:genetics / Q000378:metabolism*; D020742:rhoA GTP-Binding Protein / Q000378:metabolism
D009185:Myelin Proteins; D000070798:Nogo Proteins; D049349:Receptors, Lysosphingolipid; C000603201:Rtn4 protein, mouse; D059470:Tetraspanins; C426223:Tspan3 protein, mouse; D020742:rhoA GTP-Binding Protein
Internalization; Myelin-associated inhibitor; Nogo-A; Receptor complex; Sphingosine 1-phosphate receptor 2; Tetraspanin