Direct visualization of the microtubule lattice seam both in vitro and in vivo.
Microtubules are constructed from alpha- and beta-tubulin heterodimers that are arranged into protofilaments. Most commonly there are 13 or 14 protofilaments. A series of structural investigations using both electron microscopy and x-ray diffraction have indicated that there are two potential lattices (A and B) in which the tubulin subunits can be arranged. Electron microscopy has shown that kinesin heads, which bind only to beta-tubulin, follow a helical path with a 12-nm pitch in which subunits repeat every 8-nm axially, implying a primarily B-type lattice. However, these helical symmetry parameters are not consistent with a closed lattice and imply that there must be a discontinuity or "seam" along the microtubule. We have used quick-freeze deep-etch electron microscopy to obtain the first direct evidence for the presence of this seam in microtubules formed either in vivo or in vitro. In addition to a conventional single seam, we have also rarely found microtubules in which there is more than one seam. Overall our data indicates that microtubules have a predominantly B lattice, but that A lattice bonds between tubulin subunits are found at the seam. The cytoplasmic microtubules in mouse nerve cells also have predominantly B lattice structure and A lattice bonds at the seam. These observations have important implications for the interaction of microtubules with MAPs and with motor proteins, and for example, suggest that kinesin motors may follow a single protofilament track.
Kikkawa
M
M
Department of Anatomy and Cell Biology, Faculty of Medicine, University of Tokyo, Japan.
Ishikawa
T
T
Nakata
T
T
Wakabayashi
T
T
Hirokawa
N
N
D000818:Animals; D002531:Cerebellum / Q000648:ultrastructure; D005613:Freeze Etching / Q000379:methods; D007091:Image Processing, Computer-Assisted; D016547:Kinesins / Q000235:genetics / Q000378:metabolism / Q000648:ultrastructure; D051379:Mice; D008854:Microscopy, Electron; D008870:Microtubules / Q000648:ultrastructure*; D008958:Models, Molecular; D008961:Models, Structural; D011485:Protein Binding; D011994:Recombinant Proteins / Q000648:ultrastructure; D013552:Swine; D014404:Tubulin / Q000378:metabolism / Q000648:ultrastructure
D011994:Recombinant Proteins; D014404:Tubulin; D016547:Kinesins