Complex formation with kinesin motor domains affects the structure of microtubules

J Mol Biol. 2004 Jan 2;335(1):139-53. doi: 10.1016/j.jmb.2003.10.039.

Abstract

Microtubules are highly dynamic components of the cytoskeleton. They are important for cell movement and they are involved in a variety of transport processes together with motor proteins, such as kinesin. The exact mechanism of these transport processes is not known and so far the focus has been on structural changes within the motor domains, but not within the underlying microtubule structure. Here we investigated the interaction between kinesin and tubulin and our experimental data show that microtubules themselves are changing structure during that process. We studied unstained, vitrified samples of microtubules composed of 15 protofilaments using cryo electron microscopy and helical image analysis. 3D maps of plain microtubules and microtubules decorated with kinesin have been reconstructed to approximately 17A resolution. The alphabeta-tubulin dimer could be identified and, according to our data, alpha- and beta-tubulin adopt different conformations in plain microtubules. Significant differences were detected between maps of plain microtubules and microtubule-kinesin complexes. Most pronounced is the continuous axial inter-dimer contact in the microtubule-kinesin complex, suggesting stabilized protofilaments along the microtubule axis. It seems, that mainly structural changes within alpha-tubulin are responsible for this observation. Lateral effects are less pronounced. Following our data, we believe, that microtubules play an active role in intracellular transport processes through modulations of their core structure.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cryoelectron Microscopy
  • Imaging, Three-Dimensional
  • Kinesins / chemistry*
  • Kinesins / metabolism
  • Microtubules / chemistry*
  • Microtubules / metabolism
  • Models, Molecular
  • Molecular Motor Proteins / chemistry
  • Molecular Motor Proteins / metabolism
  • Neurospora crassa / chemistry
  • Protein Binding
  • Protein Conformation
  • Protein Subunits / chemistry
  • Tubulin / chemistry

Substances

  • Molecular Motor Proteins
  • Protein Subunits
  • Tubulin
  • Kinesins