The C-terminus of tubulin increases cytoplasmic dynein and kinesin processivity.
In motor movement on microtubules, the anionic C-terminal of tubulin has been implicated as a significant factor. Our digital analyses of movements of cytoplasmic dynein- and kinesin-coated beads on microtubules have revealed dramatic changes when the C-terminal region (2-4-kDa fragment) of tubulin was cleaved by limited subtilisin digestion of assembled microtubules. For both motors, bead binding to microtubules was decreased threefold, bead run length was decreased over fourfold, and there was a dramatic 20-fold decrease in diffusional movements of cytoplasmic dynein beads on microtubules (even with low motor concentrations where the level of bead motile activity was linear with motor concentration). The velocity of active bead movements on microtubules was unchanged for cytoplasmic dynein and slightly decreased for kinesin. There was also a decrease in the frequency of bead movements without a change in velocity when the ionic strength was raised. However, with high ionic strength there was not a decrease in run length or any selective inhibition of the diffusional movement. The C-terminal region of tubulin increased motor run length (processivity) by inhibiting "detachment" but without affecting velocity. Because the major motor binding sites of microtubules are not on the C-terminal tail of tubulin (), we suggest that the changes are the result of the compromise of a weakly attached state that is the lowest affinity step in both motors' ATPase cycles and is not rate limiting.
Wang
Z
Z
Department of Cell Biology, Duke University Medical Center, Durham, North Carolina 27710, USA.
Sheetz
M P
MP
D000818:Animals; D001665:Binding Sites; D055592:Biophysical Phenomena; D001703:Biophysics; D002642:Chick Embryo; D004398:Dyneins / Q000378:metabolism*; D066298:In Vitro Techniques; D016547:Kinesins / Q000378:metabolism*; D008870:Microtubules / Q000378:metabolism; D020409:Molecular Motor Proteins / Q000378:metabolism; D011189:Potassium Chloride; D020860:Subtilisin; D014404:Tubulin / Q000737:chemistry* / Q000378:metabolism*
D020409:Molecular Motor Proteins; D014404:Tubulin; D011189:Potassium Chloride; D020860:Subtilisin; D004398:Dyneins; D016547:Kinesins