Kidins220/ARMS Is Transported by a Kinesin-1–based Mechanism Likely to be Involved in Neuronal Differentiation Bracale, Aurora Cesca, Fabrizia Neubrand, Veronika Elisabeth Newsome, Timothy P Way, Michael Schiavo, Giampietro Kidins220/ARMS Kinesin-1 Neuronal differentiation We thank C. L. Thomas for the preparation of the KC construct and advice, K. J. Verhey for the pcDNA3-HA-KLC-HR and pcDNA3-HA-KLC-TPR plas- mids, D. Badford and J. Yang for the Pp5 construct, and V. Cavalli for the SyD/JIP-3 cDNA and antibody. We thank D. Worth for the cloning of the KLC1 constructs, A. Nicol for the Mathematica workbook used for tracking data analysis, and P. Fitzjohn for the structure prediction analysis of KIM and KLC. We are grateful to S. Tooze, A. Behrens, and members of the Molecular Neuropathobiology laboratory for critical reading of the manuscript. This work was supported by Cancer Research UK. Kinase D-interacting substrate of 220 kDa/ankyrin repeat-rich membrane spanning (Kidins220/ARMS) is a conserved membrane protein mainly expressed in brain and neuroendocrine cells, which is a downstream target of the signaling cascades initiated by neurotrophins and ephrins. We identified kinesin light chain 1 (KLC1) as a binding partner for Kidins220/ARMS by a yeast two-hybrid screen. The interaction between Kidins220/ARMS and the kinesin-1 motor complex was confirmed by glutathione S-transferase-pull-down and coimmunoprecipitation experiments. In addition, Kidins220/ARMS and kinesin-1 were shown to colocalize in nerve growth factor (NGF)-differentiated PC12 cells. Using Kidins220/ARMS and KLC1 mutants, we mapped the regions responsible for the binding to a short sequence of Kidins220/ARMS, termed KLC-interacting motif (KIM), which is sufficient for the interaction with KLC1. Optimal binding of KIM requires a region of KLC1 spanning both the tetratricopeptide repeats and the heptad repeats, previously not involved in cargo recognition. Overexpression of KIM in differentiating PC12 cells impairs the formation and transport of EGFP-Kidins220/ARMS carriers to the tips of growing neurites, leaving other kinesin-1 dependent processes unaffected. Furthermore, KIM overexpression interferes with the activation of the mitogen-activated protein kinase signaling and neurite outgrowth in NGF-treated PC12 cells. Our results suggest that Kidins220/ARMS-positive carriers undergo a kinesin-1-dependent transport linked to neurotrophin action. 2024-01-03T11:54:35Z 2024-01-03T11:54:35Z 2007 moving image Mol Biol Cell 18: 142–152 https://hdl.handle.net/10481/86549 https://doi.org/10.1091/mbc.e06-05-0453 eng http://creativecommons.org/licenses/by-nc-nd/3.0/ open access Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License The American Society for Cell Biology