The structural basis of force generation by the mitotic motor kinesin-5

Adeline Goulet, William M. Behnke-Parks, Charles V. Sindelar, Jennifer Major, Steven S. Rosenfeld, Carolyn A. Moores

Research output: Contribution to journalArticle

49 Scopus citations

Abstract

Kinesin-5 is required for forming the bipolar spindle during mitosis. Its motor domain, which contains nucleotide and microtubule binding sites and mechanical elements to generate force, has evolved distinct properties for its spindle-based functions. In this study, we report subnanometer resolution cryoelectron microscopy reconstructions of microtubule-bound human kinesin-5 before and after nucleotide binding and combine this information with studies of the kinetics of nucleotideinduced neck linker and cover strand movement. These studies reveal coupled, nucleotide-dependent conformational changes that explain many of this motor's properties. We find that ATP binding induces a ratchet-like docking of the neck linker and simultaneous, parallel docking of the N-terminal cover strand. Loop L5, the binding site for allosteric inhibitors of kinesin-5, also undergoes a dramatic reorientation when ATP binds, suggesting that it is directly involved in controlling nucleotide binding. Our structures indicate that allosteric inhibitors of human kinesin-5, which are being developed as anti-cancer therapeutics, bind to a motor conformation that occurs in the course of normal function. However, due to evolutionarily defined sequence variations in L5, this conformation is not adopted by invertebrate kinesin-5s, explaining their resistance to drug inhibition. Together, our data reveal the precision with which the molecular mechanism of kinesin-5 motors has evolved for force generation.

Original languageEnglish (US)
Pages (from-to)44654-44666
Number of pages13
JournalJournal of Biological Chemistry
Volume287
Issue number53
DOIs
StatePublished - Dec 28 2012

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ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Goulet, A., Behnke-Parks, W. M., Sindelar, C. V., Major, J., Rosenfeld, S. S., & Moores, C. A. (2012). The structural basis of force generation by the mitotic motor kinesin-5. Journal of Biological Chemistry, 287(53), 44654-44666. https://doi.org/10.1074/jbc.M112.404228