Image reconstructions of microtubules decorated with monomeric and dimeric kinesins: comparison with x-ray structure and implications for motility.
We have decorated microtubules with monomeric and dimeric kinesin constructs, studied their structure by cryoelectron microscopy and three-dimensional image reconstruction, and compared the results with the x-ray crystal structure of monomeric and dimeric kinesin. A monomeric kinesin construct (rK354, containing only a short neck helix insufficient for coiled-coil formation) decorates microtubules with a stoichiometry of one kinesin head per tubulin subunit (alpha-beta-heterodimer). The orientation of the kinesin head (an anterograde motor) on the microtubule surface is similar to that of ncd (a retrograde motor). A longer kinesin construct (rK379) forms a dimer because of the longer neck helix forming a coiled-coil. Unexpectedly, this construct also decorates the microtubule with a stoichiometry of one head per tubulin subunit, and the orientation is similar to that of the monomeric construct. This means that the interaction with microtubules causes the two heads of a kinesin dimer to separate sufficiently so that they can bind to two different tubulin subunits. This result is in contrast to recent models and can be explained by assuming that the tubulin-kinesin interaction is antagonistic to the coiled-coil interaction within a kinesin dimer.
Hoenger
A
A
Institute of Cell Biology, Swiss Federal Institute of Technology, ETH-Hönggerberg, CH-8093 Zürich, Switzerland.
Sack
S
S
Thormählen
M
M
Marx
A
A
Müller
J
J
Gross
H
H
Mandelkow
E
E
D000818:Animals; D018360:Crystallography, X-Ray; D019281:Dimerization; D007091:Image Processing, Computer-Assisted / Q000379:methods*; D016547:Kinesins / Q000737:chemistry* / Q000378:metabolism / Q000648:ultrastructure*; D008854:Microscopy, Electron / Q000379:methods; D008870:Microtubules / Q000378:metabolism / Q000648:ultrastructure*; D011487:Protein Conformation; D051381:Rats; D014404:Tubulin / Q000737:chemistry / Q000648:ultrastructure
D014404:Tubulin; D016547:Kinesins