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. 1992 Oct;3(10):1141–1154. doi: 10.1091/mbc.3.10.1141

Modulation of the dynamic instability of tubulin assembly by the microtubule-associated protein tau.

D N Drechsel 1, A A Hyman 1, M H Cobb 1, M W Kirschner 1
PMCID: PMC275678  PMID: 1421571

Abstract

Microtubule-associated proteins (MAP), such as tau, modulate the extent and rate of microtubule assembly and play an essential role in morphogenetic processes, such as axonal growth. We have examined the mechanism by which tau affects microtubule polymerization by examining the kinetics of microtubule assembly and disassembly through direct observation of microtubules using dark-field microscopy. Tau increases the rate of polymerization, decreases the rate of transit into the shrinking phase (catastrophe), and inhibits the rate of depolymerization. Tau strongly suppresses the catastrophe rate, and its ability to do so is independent of its ability to increase the elongation rate. Thus, tau generates a partially stable but still dynamic state in microtubules. This state is perturbed by phosphorylation by MAP2 kinase, which affects all three activities by lowering the affinity of tau for the microtubule lattice.

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Selected References

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  1. Allen C., Borisy G. G. Structural polarity and directional growth of microtubules of Chlamydomonas flagella. J Mol Biol. 1974 Dec 5;90(2):381–402. doi: 10.1016/0022-2836(74)90381-7. [DOI] [PubMed] [Google Scholar]
  2. Black M. M., Aletta J. M., Greene L. A. Regulation of microtubule composition and stability during nerve growth factor-promoted neurite outgrowth. J Cell Biol. 1986 Aug;103(2):545–557. doi: 10.1083/jcb.103.2.545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boulton T. G., Gregory J. S., Cobb M. H. Purification and properties of extracellular signal-regulated kinase 1, an insulin-stimulated microtubule-associated protein 2 kinase. Biochemistry. 1991 Jan 8;30(1):278–286. doi: 10.1021/bi00215a038. [DOI] [PubMed] [Google Scholar]
  4. Boulton T. G., Nye S. H., Robbins D. J., Ip N. Y., Radziejewska E., Morgenbesser S. D., DePinho R. A., Panayotatos N., Cobb M. H., Yancopoulos G. D. ERKs: a family of protein-serine/threonine kinases that are activated and tyrosine phosphorylated in response to insulin and NGF. Cell. 1991 May 17;65(4):663–675. doi: 10.1016/0092-8674(91)90098-j. [DOI] [PubMed] [Google Scholar]
  5. Bré M. H., Karsenti E. Effects of brain microtubule-associated proteins on microtubule dynamics and the nucleating activity of centrosomes. Cell Motil Cytoskeleton. 1990;15(2):88–98. doi: 10.1002/cm.970150205. [DOI] [PubMed] [Google Scholar]
  6. Butler M., Shelanski M. L. Microheterogeneity of microtubule-associated tau proteins is due to differences in phosphorylation. J Neurochem. 1986 Nov;47(5):1517–1522. doi: 10.1111/j.1471-4159.1986.tb00788.x. [DOI] [PubMed] [Google Scholar]
  7. Butner K. A., Kirschner M. W. Tau protein binds to microtubules through a flexible array of distributed weak sites. J Cell Biol. 1991 Nov;115(3):717–730. doi: 10.1083/jcb.115.3.717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Caceres A., Kosik K. S. Inhibition of neurite polarity by tau antisense oligonucleotides in primary cerebellar neurons. Nature. 1990 Feb 1;343(6257):461–463. doi: 10.1038/343461a0. [DOI] [PubMed] [Google Scholar]
  9. Chrétien D., Metoz F., Verde F., Karsenti E., Wade R. H. Lattice defects in microtubules: protofilament numbers vary within individual microtubules. J Cell Biol. 1992 Jun;117(5):1031–1040. doi: 10.1083/jcb.117.5.1031. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Cleveland D. W., Hwo S. Y., Kirschner M. W. Physical and chemical properties of purified tau factor and the role of tau in microtubule assembly. J Mol Biol. 1977 Oct 25;116(2):227–247. doi: 10.1016/0022-2836(77)90214-5. [DOI] [PubMed] [Google Scholar]
  11. Cleveland D. W., Hwo S. Y., Kirschner M. W. Purification of tau, a microtubule-associated protein that induces assembly of microtubules from purified tubulin. J Mol Biol. 1977 Oct 25;116(2):207–225. doi: 10.1016/0022-2836(77)90213-3. [DOI] [PubMed] [Google Scholar]
  12. Cobb M. H., Boulton T. G., Robbins D. J. Extracellular signal-regulated kinases: ERKs in progress. Cell Regul. 1991 Dec;2(12):965–978. doi: 10.1091/mbc.2.12.965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Daniels M. P. Colchicine inhibition of nerve fiber formation in vitro. J Cell Biol. 1972 Apr;53(1):164–176. doi: 10.1083/jcb.53.1.164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Drewes G., Lichtenberg-Kraag B., Döring F., Mandelkow E. M., Biernat J., Goris J., Dorée M., Mandelkow E. Mitogen activated protein (MAP) kinase transforms tau protein into an Alzheimer-like state. EMBO J. 1992 Jun;11(6):2131–2138. doi: 10.1002/j.1460-2075.1992.tb05272.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Drubin D. G., Feinstein S. C., Shooter E. M., Kirschner M. W. Nerve growth factor-induced neurite outgrowth in PC12 cells involves the coordinate induction of microtubule assembly and assembly-promoting factors. J Cell Biol. 1985 Nov;101(5 Pt 1):1799–1807. doi: 10.1083/jcb.101.5.1799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Evans L., Mitchison T., Kirschner M. Influence of the centrosome on the structure of nucleated microtubules. J Cell Biol. 1985 Apr;100(4):1185–1191. doi: 10.1083/jcb.100.4.1185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gildersleeve R. F., Cross A. R., Cullen K. E., Fagen A. P., Williams R. C., Jr Microtubules grow and shorten at intrinsically variable rates. J Biol Chem. 1992 Apr 25;267(12):7995–8006. [PubMed] [Google Scholar]
  18. Gotoh Y., Nishida E., Matsuda S., Shiina N., Kosako H., Shiokawa K., Akiyama T., Ohta K., Sakai H. In vitro effects on microtubule dynamics of purified Xenopus M phase-activated MAP kinase. Nature. 1991 Jan 17;349(6306):251–254. doi: 10.1038/349251a0. [DOI] [PubMed] [Google Scholar]
  19. Grundke-Iqbal I., Iqbal K., Tung Y. C., Quinlan M., Wisniewski H. M., Binder L. I. Abnormal phosphorylation of the microtubule-associated protein tau (tau) in Alzheimer cytoskeletal pathology. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4913–4917. doi: 10.1073/pnas.83.13.4913. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Haycock J. W., Ahn N. G., Cobb M. H., Krebs E. G. ERK1 and ERK2, two microtubule-associated protein 2 kinases, mediate the phosphorylation of tyrosine hydroxylase at serine-31 in situ. Proc Natl Acad Sci U S A. 1992 Mar 15;89(6):2365–2369. doi: 10.1073/pnas.89.6.2365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hilbush B. S., Levine J. M. Stimulation of a Ca(2+)-dependent protein kinase by GM1 ganglioside in nerve growth factor-treated PC12 cells. Proc Natl Acad Sci U S A. 1991 Jul 1;88(13):5616–5620. doi: 10.1073/pnas.88.13.5616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Hill T. L., Carlier M. F. Steady-state theory of the interference of GTP hydrolysis in the mechanism of microtubule assembly. Proc Natl Acad Sci U S A. 1983 Dec;80(23):7234–7238. doi: 10.1073/pnas.80.23.7234. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Himmler A., Drechsel D., Kirschner M. W., Martin D. W., Jr Tau consists of a set of proteins with repeated C-terminal microtubule-binding domains and variable N-terminal domains. Mol Cell Biol. 1989 Apr;9(4):1381–1388. doi: 10.1128/mcb.9.4.1381. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Horio T., Hotani H. Visualization of the dynamic instability of individual microtubules by dark-field microscopy. Nature. 1986 Jun 5;321(6070):605–607. doi: 10.1038/321605a0. [DOI] [PubMed] [Google Scholar]
  25. Hyman A. A. Preparation of marked microtubules for the assay of the polarity of microtubule-based motors by fluorescence. J Cell Sci Suppl. 1991;14:125–127. doi: 10.1242/jcs.1991.supplement_14.25. [DOI] [PubMed] [Google Scholar]
  26. Hyman A. A., Salser S., Drechsel D. N., Unwin N., Mitchison T. J. Role of GTP hydrolysis in microtubule dynamics: information from a slowly hydrolyzable analogue, GMPCPP. Mol Biol Cell. 1992 Oct;3(10):1155–1167. doi: 10.1091/mbc.3.10.1155. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Hyman A., Drechsel D., Kellogg D., Salser S., Sawin K., Steffen P., Wordeman L., Mitchison T. Preparation of modified tubulins. Methods Enzymol. 1991;196:478–485. doi: 10.1016/0076-6879(91)96041-o. [DOI] [PubMed] [Google Scholar]
  28. Kirschner M., Mitchison T. Beyond self-assembly: from microtubules to morphogenesis. Cell. 1986 May 9;45(3):329–342. doi: 10.1016/0092-8674(86)90318-1. [DOI] [PubMed] [Google Scholar]
  29. Knops J., Kosik K. S., Lee G., Pardee J. D., Cohen-Gould L., McConlogue L. Overexpression of tau in a nonneuronal cell induces long cellular processes. J Cell Biol. 1991 Aug;114(4):725–733. doi: 10.1083/jcb.114.4.725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Koshland D. E., Mitchison T. J., Kirschner M. W. Polewards chromosome movement driven by microtubule depolymerization in vitro. Nature. 1988 Feb 11;331(6156):499–504. doi: 10.1038/331499a0. [DOI] [PubMed] [Google Scholar]
  31. Landreth G. E., Smith D. S., McCabe C., Gittinger C. Characterization of a nerve growth factor-stimulated protein kinase in PC12 cells which phosphorylates microtubule-associated protein 2 and pp250. J Neurochem. 1990 Aug;55(2):514–523. doi: 10.1111/j.1471-4159.1990.tb04165.x. [DOI] [PubMed] [Google Scholar]
  32. Lichtenberg-Kraag B., Mandelkow E. M., Biernat J., Steiner B., Schröter C., Gustke N., Meyer H. E., Mandelkow E. Phosphorylation-dependent epitopes of neurofilament antibodies on tau protein and relationship with Alzheimer tau. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5384–5388. doi: 10.1073/pnas.89.12.5384. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Lindwall G., Cole R. D. Phosphorylation affects the ability of tau protein to promote microtubule assembly. J Biol Chem. 1984 Apr 25;259(8):5301–5305. [PubMed] [Google Scholar]
  34. Lindwall G., Cole R. D. The purification of tau protein and the occurrence of two phosphorylation states of tau in brain. J Biol Chem. 1984 Oct 10;259(19):12241–12245. [PubMed] [Google Scholar]
  35. Mandelkow E. M., Mandelkow E., Milligan R. A. Microtubule dynamics and microtubule caps: a time-resolved cryo-electron microscopy study. J Cell Biol. 1991 Sep;114(5):977–991. doi: 10.1083/jcb.114.5.977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Mitchison T., Kirschner M. Dynamic instability of microtubule growth. Nature. 1984 Nov 15;312(5991):237–242. doi: 10.1038/312237a0. [DOI] [PubMed] [Google Scholar]
  37. Mitchison T., Kirschner M. Microtubule assembly nucleated by isolated centrosomes. Nature. 1984 Nov 15;312(5991):232–237. doi: 10.1038/312232a0. [DOI] [PubMed] [Google Scholar]
  38. Murphy D. B., Johnson K. A., Borisy G. G. Role of tubulin-associated proteins in microtubule nucleation and elongation. J Mol Biol. 1977 Nov 25;117(1):33–52. doi: 10.1016/0022-2836(77)90021-3. [DOI] [PubMed] [Google Scholar]
  39. Nose P. S., Griffith L. C., Schulman H. Ca2+-dependent phosphorylation of tyrosine hydroxylase in PC12 cells. J Cell Biol. 1985 Oct;101(4):1182–1190. doi: 10.1083/jcb.101.4.1182. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. O'Brien E. T., Salmon E. D., Walker R. A., Erickson H. P. Effects of magnesium on the dynamic instability of individual microtubules. Biochemistry. 1990 Jul 17;29(28):6648–6656. doi: 10.1021/bi00480a014. [DOI] [PubMed] [Google Scholar]
  41. O'Brien E. T., Voter W. A., Erickson H. P. GTP hydrolysis during microtubule assembly. Biochemistry. 1987 Jun 30;26(13):4148–4156. doi: 10.1021/bi00387a061. [DOI] [PubMed] [Google Scholar]
  42. Savage C., Hamelin M., Culotti J. G., Coulson A., Albertson D. G., Chalfie M. mec-7 is a beta-tubulin gene required for the production of 15-protofilament microtubules in Caenorhabditis elegans. Genes Dev. 1989 Jun;3(6):870–881. doi: 10.1101/gad.3.6.870. [DOI] [PubMed] [Google Scholar]
  43. Schilstra M. J., Martin S. R., Bayley P. M. On the relationship between nucleotide hydrolysis and microtubule assembly: studies with a GTP-regenerating system. Biochem Biophys Res Commun. 1987 Sep 15;147(2):588–595. doi: 10.1016/0006-291x(87)90971-5. [DOI] [PubMed] [Google Scholar]
  44. Schulze E., Kirschner M. Microtubule dynamics in interphase cells. J Cell Biol. 1986 Mar;102(3):1020–1031. doi: 10.1083/jcb.102.3.1020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Steiner B., Mandelkow E. M., Biernat J., Gustke N., Meyer H. E., Schmidt B., Mieskes G., Söling H. D., Drechsel D., Kirschner M. W. Phosphorylation of microtubule-associated protein tau: identification of the site for Ca2(+)-calmodulin dependent kinase and relationship with tau phosphorylation in Alzheimer tangles. EMBO J. 1990 Nov;9(11):3539–3544. doi: 10.1002/j.1460-2075.1990.tb07563.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Stewart R. J., Farrell K. W., Wilson L. Role of GTP hydrolysis in microtubule polymerization: evidence for a coupled hydrolysis mechanism. Biochemistry. 1990 Jul 10;29(27):6489–6498. doi: 10.1021/bi00479a022. [DOI] [PubMed] [Google Scholar]
  47. Studier F. W., Rosenberg A. H., Dunn J. J., Dubendorff J. W. Use of T7 RNA polymerase to direct expression of cloned genes. Methods Enzymol. 1990;185:60–89. doi: 10.1016/0076-6879(90)85008-c. [DOI] [PubMed] [Google Scholar]
  48. Summers K., Kirschner M. W. Characteristics of the polar assembly and disassembly of microtubules observed in vitro by darkfield light microscopy. J Cell Biol. 1979 Oct;83(1):205–217. doi: 10.1083/jcb.83.1.205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Tsao H., Aletta J. M., Greene L. A. Nerve growth factor and fibroblast growth factor selectively activate a protein kinase that phosphorylates high molecular weight microtubule-associated proteins. Detection, partial purification, and characterization in PC12 cells. J Biol Chem. 1990 Sep 15;265(26):15471–15480. [PubMed] [Google Scholar]
  50. Uéda K., Masliah E., Saitoh T., Bakalis S. L., Scoble H., Kosik K. S. Alz-50 recognizes a phosphorylated epitope of tau protein. J Neurosci. 1990 Oct;10(10):3295–3304. doi: 10.1523/JNEUROSCI.10-10-03295.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. Walker R. A., O'Brien E. T., Pryer N. K., Soboeiro M. F., Voter W. A., Erickson H. P., Salmon E. D. Dynamic instability of individual microtubules analyzed by video light microscopy: rate constants and transition frequencies. J Cell Biol. 1988 Oct;107(4):1437–1448. doi: 10.1083/jcb.107.4.1437. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Walker R. A., Pryer N. K., Salmon E. D. Dilution of individual microtubules observed in real time in vitro: evidence that cap size is small and independent of elongation rate. J Cell Biol. 1991 Jul;114(1):73–81. doi: 10.1083/jcb.114.1.73. [DOI] [PMC free article] [PubMed] [Google Scholar]
  53. Weingarten M. D., Lockwood A. H., Hwo S. Y., Kirschner M. W. A protein factor essential for microtubule assembly. Proc Natl Acad Sci U S A. 1975 May;72(5):1858–1862. doi: 10.1073/pnas.72.5.1858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  54. Yamada K. M., Spooner B. S., Wessells N. K. Axon growth: roles of microfilaments and microtubules. Proc Natl Acad Sci U S A. 1970 Aug;66(4):1206–1212. doi: 10.1073/pnas.66.4.1206. [DOI] [PMC free article] [PubMed] [Google Scholar]

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