A Role for Kif17 in Transport of Kv4.2

Po-Ju Chu,Jacqueline F Rivera,Don B Arnold

doi:10.1074/jbc.m508897200

Po-Ju Chu, Jacqueline F Rivera + Show 1 more

Open Access

https://doi.org/10.1074/jbc.m508897200

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Abstract

Although kinesins are known to transport neuronal proteins, it is not known what role they play in the targeting of their cargos to specific subcellular compartments in neurons. Here we present evidence that the K+ channel Kv4.2, which is a major regulator of dendritic excitability, is transported to dendrites by the kinesin isoform Kif17. We show that a dominant negative construct against Kif17 dramatically inhibits localization to dendrites of both introduced and endogenous Kv4.2, but those against other kinesins found in dendrites do not. Kv4.2 colocalizes with Kif17 but not with other kinesin isoforms in dendrites of cortical neurons. Native Kv4.2 and Kif17 coimmunoprecipitate from brain lysate, and introduced, tagged versions of the two proteins coimmunoprecipitate from COS cell lysate, indicating that the two proteins interact, either directly or indirectly. The interaction between Kif17 and Kv4.2 appears to occur through the extreme C terminus of Kv4.2 and not through the dileucine motif. Thus, the dileucine motif does not determine the localization of Kv4.2 by causing the channel to interact with a specific motor protein. In support of this conclusion, we found that the dileucine motif mediates dendritic targeting of CD8 independent of Kif17. Together our data show that Kif17 is probably the motor that transports Kv4.2 to dendrites but suggest that this motor does not, by itself, specify dendritic localization of the channel.

Highlights

Progress toward elucidating the molecular mechanisms underlying subcellular localization of neuronal proteins has been made by identifying targeting motifs within the primary structure of transmembrane proteins that are localized in distinct subcellular compartments (4 –9)
DNKifs under the control of a constitutively active cytomegalovirus promoter were expressed in cortical neurons, and 24 h subsequently, Kv4.2-MYC was expressed in the same cells using an inducible ecdysone promoter (Fig. 1B)
We present evidence that the voltage-gated Kϩ channel Kv4.2 is transported by the kinesin isoform Kif17 in cortical neurons

Summary

Introduction

Progress toward elucidating the molecular mechanisms underlying subcellular localization of neuronal proteins has been made by identifying targeting motifs within the primary structure of transmembrane proteins that are localized in distinct subcellular compartments (4 –9). In order to quantify the effect of particular DNKifs on localization of Kv4.2, we calculated the ratio of the expression level of the tagged channel in the dendrites versus that in the cell body. In cells expressing GFP-DNKif17, the dendrite/cell body ratio of Kv4.2MYC was 0.06 Ϯ 0.003 (n ϭ 25; Fig. 1D), indicating that the channel was

Results

Conclusion