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. 1994 Dec 2;127(6):1965–1971. doi: 10.1083/jcb.127.6.1965

Direct visualization of the microtubule lattice seam both in vitro and in vivo

PMCID: PMC2120284  PMID: 7806574

Abstract

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.

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Selected References

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  1. Amos L., Klug A. Arrangement of subunits in flagellar microtubules. J Cell Sci. 1974 May;14(3):523–549. doi: 10.1242/jcs.14.3.523. [DOI] [PubMed] [Google Scholar]
  2. Beese L., Stubbs G., Cohen C. Microtubule structure at 18 A resolution. J Mol Biol. 1987 Mar 20;194(2):257–264. doi: 10.1016/0022-2836(87)90373-1. [DOI] [PubMed] [Google Scholar]
  3. Chrétien D., Wade R. H. New data on the microtubule surface lattice. Biol Cell. 1991;71(1-2):161–174. doi: 10.1016/0248-4900(91)90062-r. [DOI] [PubMed] [Google Scholar]
  4. Erickson H. P. Microtubule surface lattice and subunit structure and observations on reassembly. J Cell Biol. 1974 Jan;60(1):153–167. doi: 10.1083/jcb.60.1.153. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Harrison B. C., Marchese-Ragona S. P., Gilbert S. P., Cheng N., Steven A. C., Johnson K. A. Decoration of the microtubule surface by one kinesin head per tubulin heterodimer. Nature. 1993 Mar 4;362(6415):73–75. doi: 10.1038/362073a0. [DOI] [PubMed] [Google Scholar]
  6. Herzog W., Weber K. Fractionation of brain microtubule-associated proteins. Isolation of two different proteins which stimulate tubulin polymerization in vitro. Eur J Biochem. 1978 Dec 1;92(1):1–8. doi: 10.1111/j.1432-1033.1978.tb12716.x. [DOI] [PubMed] [Google Scholar]
  7. Heuser J. E., Salpeter S. R. Organization of acetylcholine receptors in quick-frozen, deep-etched, and rotary-replicated Torpedo postsynaptic membrane. J Cell Biol. 1979 Jul;82(1):150–173. doi: 10.1083/jcb.82.1.150. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hirokawa N. Cross-linker system between neurofilaments, microtubules, and membranous organelles in frog axons revealed by the quick-freeze, deep-etching method. J Cell Biol. 1982 Jul;94(1):129–142. doi: 10.1083/jcb.94.1.129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hirokawa N., Pfister K. K., Yorifuji H., Wagner M. C., Brady S. T., Bloom G. S. Submolecular domains of bovine brain kinesin identified by electron microscopy and monoclonal antibody decoration. Cell. 1989 Mar 10;56(5):867–878. doi: 10.1016/0092-8674(89)90691-0. [DOI] [PubMed] [Google Scholar]
  10. Hirokawa N., Shiomura Y., Okabe S. Tau proteins: the molecular structure and mode of binding on microtubules. J Cell Biol. 1988 Oct;107(4):1449–1459. doi: 10.1083/jcb.107.4.1449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Huang T. G., Suhan J., Hackney D. D. Drosophila kinesin motor domain extending to amino acid position 392 is dimeric when expressed in Escherichia coli. J Biol Chem. 1994 Jun 10;269(23):16502–16507. [PubMed] [Google Scholar]
  12. Kato Kikuya. A Collection of cDNA Clones with Specific Expression Patterns in Mouse Brain. Eur J Neurosci. 1990;2(8):704–711. doi: 10.1111/j.1460-9568.1990.tb00460.x. [DOI] [PubMed] [Google Scholar]
  13. Linck R. W. Flagellar doublet microtubules: fractionation of minor components and alpha-tubulin from specific regions of the A-tubule. J Cell Sci. 1976 Mar;20(2):405–439. doi: 10.1242/jcs.20.2.405. [DOI] [PubMed] [Google Scholar]
  14. Linck R. W., Olson G. E., Langevin G. L. Arrangement of tubulin subunits and microtubule-associated proteins in the central-pair microtubule apparatus of squid (Loligo pealei) sperm flagella. J Cell Biol. 1981 May;89(2):309–322. doi: 10.1083/jcb.89.2.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Mandelkow E. M., Schultheiss R., Rapp R., Müller M., Mandelkow E. On the surface lattice of microtubules: helix starts, protofilament number, seam, and handedness. J Cell Biol. 1986 Mar;102(3):1067–1073. doi: 10.1083/jcb.102.3.1067. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. McEwen B., Edelstein S. J. Evidence for a mixed lattice in microtubules reassembled in vitro. J Mol Biol. 1980 May 15;139(2):123–145. doi: 10.1016/0022-2836(80)90300-9. [DOI] [PubMed] [Google Scholar]
  17. Ray S., Meyhöfer E., Milligan R. A., Howard J. Kinesin follows the microtubule's protofilament axis. J Cell Biol. 1993 Jun;121(5):1083–1093. doi: 10.1083/jcb.121.5.1083. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Romberg L., Vale R. D. Chemomechanical cycle of kinesin differs from that of myosin. Nature. 1993 Jan 14;361(6408):168–170. doi: 10.1038/361168a0. [DOI] [PubMed] [Google Scholar]
  19. Scholey J. M., Heuser J., Yang J. T., Goldstein L. S. Identification of globular mechanochemical heads of kinesin. Nature. 1989 Mar 23;338(6213):355–357. doi: 10.1038/338355a0. [DOI] [PubMed] [Google Scholar]
  20. Shelanski M. L., Gaskin F., Cantor C. R. Microtubule assembly in the absence of added nucleotides. Proc Natl Acad Sci U S A. 1973 Mar;70(3):765–768. doi: 10.1073/pnas.70.3.765. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Song Y. H., Mandelkow E. Recombinant kinesin motor domain binds to beta-tubulin and decorates microtubules with a B surface lattice. Proc Natl Acad Sci U S A. 1993 Mar 1;90(5):1671–1675. doi: 10.1073/pnas.90.5.1671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Vale R. D., Reese T. S., Sheetz M. P. Identification of a novel force-generating protein, kinesin, involved in microtubule-based motility. Cell. 1985 Aug;42(1):39–50. doi: 10.1016/s0092-8674(85)80099-4. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Vale R. D., Toyoshima Y. Y. Rotation and translocation of microtubules in vitro induced by dyneins from Tetrahymena cilia. Cell. 1988 Feb 12;52(3):459–469. doi: 10.1016/s0092-8674(88)80038-2. [DOI] [PubMed] [Google Scholar]
  24. Wakabayashi T., Kubota M., Yoshida M., Kagawa Y. Structure of ATPase (coupling factor TF1) from a thermophilic bacterium. J Mol Biol. 1977 Dec 5;117(2):515–519. doi: 10.1016/0022-2836(77)90140-1. [DOI] [PubMed] [Google Scholar]
  25. Yang J. T., Saxton W. M., Stewart R. J., Raff E. C., Goldstein L. S. Evidence that the head of kinesin is sufficient for force generation and motility in vitro. Science. 1990 Jul 6;249(4964):42–47. doi: 10.1126/science.2142332. [DOI] [PubMed] [Google Scholar]

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