The bipolar mitotic kinesin Eg5 moves on both microtubules that it crosslinks.
During cell division, mitotic spindles are assembled by microtubule-based motor proteins. The bipolar organization of spindles is essential for proper segregation of chromosomes, and requires plus-end-directed homotetrameric motor proteins of the widely conserved kinesin-5 (BimC) family. Hypotheses for bipolar spindle formation include the 'push-pull mitotic muscle' model, in which kinesin-5 and opposing motor proteins act between overlapping microtubules. However, the precise roles of kinesin-5 during this process are unknown. Here we show that the vertebrate kinesin-5 Eg5 drives the sliding of microtubules depending on their relative orientation. We found in controlled in vitro assays that Eg5 has the remarkable capability of simultaneously moving at approximately 20 nm s(-1) towards the plus-ends of each of the two microtubules it crosslinks. For anti-parallel microtubules, this results in relative sliding at approximately 40 nm s(-1), comparable to spindle pole separation rates in vivo. Furthermore, we found that Eg5 can tether microtubule plus-ends, suggesting an additional microtubule-binding mode for Eg5. Our results demonstrate how members of the kinesin-5 family are likely to function in mitosis, pushing apart interpolar microtubules as well as recruiting microtubules into bundles that are subsequently polarized by relative sliding.
Kapitein
Lukas C
LC
Department of Physics and Astronomy and Laser Centre, Vrije Universiteit, De Boelelaan 1081, 1081 HV Amsterdam, The Netherlands.
Peterman
Erwin J G
EJ
Kwok
Benjamin H
BH
Kim
Jeffrey H
JH
Kapoor
Tarun M
TM
Schmidt
Christoph F
CF
D000818:Animals; D003432:Cross-Linking Reagents / Q000378:metabolism; D004058:Diffusion; D016547:Kinesins / Q000378:metabolism*; D008870:Microtubules / Q000737:chemistry / Q000378:metabolism*; D008938:Mitosis; D008954:Models, Biological; D009068:Movement; D011485:Protein Binding; D008941:Spindle Apparatus / Q000737:chemistry / Q000378:metabolism; D014981:Xenopus / Q000378:metabolism*; D029867:Xenopus Proteins / Q000378:metabolism*
D003432:Cross-Linking Reagents; C088335:KIF11 protein, Xenopus; D029867:Xenopus Proteins; D016547:Kinesins