Association of GFP-KIF17^G754E with MT plus ends requires kinesin ATPase and MT-binding activities. (A) Single frame from a time-lapse recording of MDCK cells expressing GFP-KIF17-FL. Inset shows a kymograph of a moving GFP-KIF17-FL puncta during 20 s of the time-lapse recording (Video 1). The inset shows an enlargement of the boxed region. (B) Alignment of the hinge regions of C. elegans OSM-3 and human KIF17. The gray box shows the position of the G-to-E mutations in KIF17 and OSM-3. Numbers at left and right indicate amino acid position. Asterisks indicate sequence identities between KIF17 and OSM-3. (C) Single frame from a time-lapse recording of Caco2 cells injected with GFP-KIF17^G754E cDNA (Video 2). Boxed regions (a and b) are enlarged in the bottom panels. (B, right) Selected frames from the time-lapse recording. Asterisks indicate a KIF17 puncta moving on MTs toward the plus end. (D) Images of GFP-KIF17^G754E, GFP-KIF17^G754E-G243A, and GFP-KIF17^{G754E-R288/294A} expressed by cDNA injection in live or prepermeabilized and fixed MDCK cells. GFP-KIF17^G754E localizes in cell extensions at MT ends (arrows), whereas GFP-KIF17^G754E-G243A and GFP-KIF17^{G754E-R288/294A} do not (arrowheads).