Abstract
Synaptic proteins are synthesized in the cell body and transported down the axon by microtubule-dependent motors. We previously reported that KIF1Bβ and KIF1A motors are essential for transporting synaptic vesicle precursors; however the mechanisms that regulate transport, as well as cargo recognition and control of cargo loading and unloading remain largely unknown. Here, we show that DENN/MADD (Rab3-GEP) is an essential part of the regulation mechanism through direct interaction with the stalk domain of KIF1Bβ and KIF1A. We also show that DENN/MADD binds preferentially to GTP–Rab3 and acts as a Rab3 effector. These molecular interactions are fundamental as sequential genetic perturbations revealed that KIF1Bβ and KIF1A are essential for the transport of DENN/MADD and Rab3, whereas DENN/MADD is essential for the transport of Rab3. GTP–Rab3 was more effectively transported than GDP–Rab3, suggesting that the nucleotide state of Rab3 regulates axonal transport of Rab3-carrying vesicles through preferential interaction with DENN/MADD.
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Acknowledgements
The authors thank H. Fukuda, H. Sato, T. Aizawa and others from the Hirokawa laboratory for discussions and technical assistance. We are also grateful to the Kazusa DNA Institute (Chiba, Japan) for providing KIAA clones. This study was supported by a Grant-in-Aid for Specially Promoted Research to N.H. and Global COE programme to University of Tokyo from the Ministry of Education, Culture, Sports, Science and Technology of Japan.
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Niwa, S., Tanaka, Y. & Hirokawa, N. KIF1Bβ- and KIF1A-mediated axonal transport of presynaptic regulator Rab3 occurs in a GTP-dependent manner through DENN/MADD. Nat Cell Biol 10, 1269–1279 (2008). https://doi.org/10.1038/ncb1785
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DOI: https://doi.org/10.1038/ncb1785
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