Insights into how JIP3 and JIP4 adaptor proteins relieve the auto-inhibited form of the kinesin-1 motor
Abstract
While our understanding of kinesin autoinhibition mechanisms is advancing, key insights into kinesin activation, especially autoinhibition relieved by cargo-motor adaptors, remain unclear. JIP3 and JIP4 are adaptors that activate kinesin-1 and dynein-dynactin motors, enabling bidirectional transport along microtubules. Here, we characterized the interaction between the kinesin-1 heavy chain, KIF5B, and both JIP3 and JIP4 (JIP3/4), using mutagenesis and binding experiments associated with modelling. We identified the minimal regions of JIP3/4 and KIF5B required for interaction. JIP3/4 binds to the CC4 coiled-coil domain of KIF5B, with no significant interaction detected with the distal tail. Conversely, KIF5B- CC4 binds to the RH1 domain of JIP3/4, with the preceding disordered N-terminus modulating the force of the binding. Notably, JIP3 and JIP4 exhibit 40-fold differences in affinity for KIF5B. We performed extensive site-directed mutagenesis on KIF5B-binding regions of JIP3/JIP4 targeting accessible charged residues and sequence differences between JIP3 and JIP4. This revealed multiple weak-affinity binding sites that collectively define an extensive KIF5B-binding surface on the RH1 domain of JIP3. Importantly, the KIF5B-binding surface on JIP3 differs from that of the DLIC (dynein light intermediate chain), confirming no competition between the two motors. Altogether, the fragment and mutant mapping of KIF5B binding to JIP3 and JIP4 presented here provides insights into how the auto-inhibited form of kinesin-1 might be relieved, with the CC4 region playing a central role.
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BibTeX
@article{vilela2026insights,
title = {{Insights into how JIP3 and JIP4 adaptor proteins relieve the auto-inhibited form of the kinesin-1 motor}},
author = {Fernando Vilela and Mélanie Chenon and Valérie Campanacci and Ania Alik and Christophe Velours and Jessica Andreani and Paola Llinas and Julie Ménétrey},
journal = {Journal of Biological Chemistry},
year = {2026},
month = jul,
doi = {10.1016/j.jbc.2026.113355},
url = {https://hal.science/hal-05767187v1},
note = {CC BY 4.0},
}
RIS
TY - JOUR
TI - Insights into how JIP3 and JIP4 adaptor proteins relieve the auto-inhibited form of the kinesin-1 motor
AU - Fernando Vilela
AU - Mélanie Chenon
AU - Valérie Campanacci
AU - Ania Alik
AU - Christophe Velours
AU - Jessica Andreani
AU - Paola Llinas
AU - Julie Ménétrey
PY - 2026
DA - 2026/07/22
JO - Journal of Biological Chemistry
DO - 10.1016/j.jbc.2026.113355
UR - https://hal.science/hal-05767187v1
AB - While our understanding of kinesin autoinhibition mechanisms is advancing, key insights into kinesin activation, especially autoinhibition relieved by cargo-motor adaptors, remain unclear. JIP3 and JIP4 are adaptors that activate kinesin-1 and dynein-dynactin motors, enabling bidirectional transport along microtubules. Here, we characterized the interaction between the kinesin-1 heavy chain, KIF5B, and both JIP3 and JIP4 (JIP3/4), using mutagenesis and binding experiments associated with modelling. We identified the minimal regions of JIP3/4 and KIF5B required for interaction. JIP3/4 binds to the CC4 coiled-coil domain of KIF5B, with no significant interaction detected with the distal tail. Conversely, KIF5B- CC4 binds to the RH1 domain of JIP3/4, with the preceding disordered N-terminus modulating the force of the binding. Notably, JIP3 and JIP4 exhibit 40-fold differences in affinity for KIF5B. We performed extensive site-directed mutagenesis on KIF5B-binding regions of JIP3/JIP4 targeting accessible charged residues and sequence differences between JIP3 and JIP4. This revealed multiple weak-affinity binding sites that collectively define an extensive KIF5B-binding surface on the RH1 domain of JIP3. Importantly, the KIF5B-binding surface on JIP3 differs from that of the DLIC (dynein light intermediate chain), confirming no competition between the two motors. Altogether, the fragment and mutant mapping of KIF5B binding to JIP3 and JIP4 presented here provides insights into how the auto-inhibited form of kinesin-1 might be relieved, with the CC4 region playing a central role.
SN - 0021-9258
SN - 1083-351X
ER -