The effect of monastrol on the processive motility of a dimeric kinesin-5 head/kinesin-1 stalk chimera.

Published

Journal Article

Controlled activity of several kinesin motors is required for the proper assembly of the mitotic spindle. Eg5, a homotetrameric bipolar kinesin-5 from Xenopus laevis, can cross-link and slide anti-parallel microtubules apart by a motility mechanism comprising diffusional and directional modes. How this mechanism is regulated, possibly by the tail domains of the opposing motors, is poorly understood. In order to explore the basic unregulated kinesin-5 motor activity, we generated a stably dimeric kinesin-5 construct, Eg5Kin, consisting of the motor domain and neck linker of Eg5 and the neck coiled coil of Drosophila melanogaster kinesin-1 (DmKHC). In single-molecule motility assays, we found this chimera to be highly processive. In addition, we studied the effect of the kinesin-5-specific inhibitor monastrol using single-molecule fluorescence assays. We found that monastrol reduced the length of processive runs, but strikingly did not affect velocity. Quantitative analysis of monastrol dose dependence suggests that two bound monastrol molecules are required to be bound to an Eg5Kin dimer to terminate a run.

Full Text

Duke Authors

Cited Authors

  • Lakämper, S; Thiede, C; Düselder, A; Reiter, S; Korneev, MJ; Kapitein, LC; Peterman, EJG; Schmidt, CF

Published Date

  • May 2010

Published In

Volume / Issue

  • 399 / 1

Start / End Page

  • 1 - 8

PubMed ID

  • 20227420

Pubmed Central ID

  • 20227420

Electronic International Standard Serial Number (EISSN)

  • 1089-8638

International Standard Serial Number (ISSN)

  • 0022-2836

Digital Object Identifier (DOI)

  • 10.1016/j.jmb.2010.03.009

Language

  • eng