Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1987 Sep 1;105(3):1273–1282. doi: 10.1083/jcb.105.3.1273

MAP 1C is a microtubule-activated ATPase which translocates microtubules in vitro and has dynein-like properties

PMCID: PMC2114794  PMID: 2958482

Abstract

We observe that one of the high molecular mass microtubule-associated proteins (MAPs) from brain exhibits nucleotide-dependent binding to microtubules. We identify the protein as MAP IC, which was previously described in this laboratory as a minor component of standard microtubule preparations (Bloom, G.S., T. Schoenfeld, and R.B. Vallee, 1984, J. Cell Biol., 98:320-330). We find that MAP 1C is enriched in microtubules prepared in the absence of nucleotide. Kinesin is also found in these preparations, but can be specifically extracted with GTP. A fraction highly enriched in MAP 1C can be prepared by subsequent extraction of the microtubules with ATP. Two activities cofractionate with MAP 1C upon further purification, a microtubule-activated ATPase activity and a microtubule-translocating activity. These activities indicate a role for the protein in cytoplasmic motility. MAP 1C coelectrophoreses with the beta heavy chain of Chlamydomonas flagellar dynein, and has a sedimentation coefficient of 20S. Exposure to ultraviolet light in the presence of vanadate and ATP results in the production of two large fragments of MAP 1C. These characteristics suggest that MAP 1C may be a cytoplasmic analogue of axonemal dynein.

Full Text

The Full Text of this article is available as a PDF (2.7 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allen R. D., Metuzals J., Tasaki I., Brady S. T., Gilbert S. P. Fast axonal transport in squid giant axon. Science. 1982 Dec 10;218(4577):1127–1129. doi: 10.1126/science.6183744. [DOI] [PubMed] [Google Scholar]
  2. Asai D. J., Wilson L. A latent activity dynein-like cytoplasmic magnesium adenosine triphosphatase. J Biol Chem. 1985 Jan 25;260(2):699–702. [PubMed] [Google Scholar]
  3. Bell C. W., Fraser C. L., Sale W. S., Tang W. J., Gibbons I. R. Preparation and purification of dynein. Methods Enzymol. 1982;85(Pt B):450–474. doi: 10.1016/0076-6879(82)85045-3. [DOI] [PubMed] [Google Scholar]
  4. Bloom G. S., Luca F. C., Vallee R. B. Microtubule-associated protein 1B: identification of a major component of the neuronal cytoskeleton. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5404–5408. doi: 10.1073/pnas.82.16.5404. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bloom G. S., Schoenfeld T. A., Vallee R. B. Widespread distribution of the major polypeptide component of MAP 1 (microtubule-associated protein 1) in the nervous system. J Cell Biol. 1984 Jan;98(1):320–330. doi: 10.1083/jcb.98.1.320. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Brady S. T. A novel brain ATPase with properties expected for the fast axonal transport motor. Nature. 1985 Sep 5;317(6032):73–75. doi: 10.1038/317073a0. [DOI] [PubMed] [Google Scholar]
  7. Breuer A. C., Christian C. N., Henkart M., Nelson P. G. Computer analysis of organelle translocation in primary neuronal cultures and continuous cell lines. J Cell Biol. 1975 Jun;65(3):562–576. doi: 10.1083/jcb.65.3.562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Burns R. G., Pollard T. D. A dynein-like protein from brain. FEBS Lett. 1974 Apr 1;40(2):274–280. doi: 10.1016/0014-5793(74)80243-7. [DOI] [PubMed] [Google Scholar]
  9. 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]
  10. Collins C. A., Vallee R. B. A microtubule-activated ATPase from sea urchin eggs, distinct from cytoplasmic dynein and kinesin. Proc Natl Acad Sci U S A. 1986 Jul;83(13):4799–4803. doi: 10.1073/pnas.83.13.4799. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Collins C. A., Vallee R. B. Characterization of the sea-urchin egg microtubule-activated ATPase. J Cell Sci Suppl. 1986;5:197–204. doi: 10.1242/jcs.1986.supplement_5.13. [DOI] [PubMed] [Google Scholar]
  12. Gaskin F., Kramer S. B., Cantor C. R., Adelstein R., Shelanski M. L. A dynein-like protein associated with neurotubules. FEBS Lett. 1974 Apr 1;40(2):281–286. doi: 10.1016/0014-5793(74)80244-9. [DOI] [PubMed] [Google Scholar]
  13. Gibbons I. R., Lee-Eiford A., Mocz G., Phillipson C. A., Tang W. J., Gibbons B. H. Photosensitized cleavage of dynein heavy chains. Cleavage at the "V1 site" by irradiation at 365 nm in the presence of ATP and vanadate. J Biol Chem. 1987 Feb 25;262(6):2780–2786. [PubMed] [Google Scholar]
  14. Gilbert S. P., Sloboda R. D. Identification of a MAP 2-like ATP-binding protein associated with axoplasmic vesicles that translocate on isolated microtubules. J Cell Biol. 1986 Sep;103(3):947–956. doi: 10.1083/jcb.103.3.947. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Herrmann H., Dalton J. M., Wiche G. Microheterogeneity of microtubule-associated proteins, MAP-1 and MAP-2, and differential phosphorylation of individual subcomponents. J Biol Chem. 1985 May 10;260(9):5797–5803. [PubMed] [Google Scholar]
  16. Hollenbeck P. J., Chapman K. A novel microtubule-associated protein from mammalian nerve shows ATP-sensitive binding to microtubules. J Cell Biol. 1986 Oct;103(4):1539–1545. doi: 10.1083/jcb.103.4.1539. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hollenbeck P. J., Suprynowicz F., Cande W. Z. Cytoplasmic dynein-like ATPase cross-links microtubules in an ATP-sensitive manner. J Cell Biol. 1984 Oct;99(4 Pt 1):1251–1258. doi: 10.1083/jcb.99.4.1251. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kim H., Binder L. I., Rosenbaum J. L. The periodic association of MAP2 with brain microtubules in vitro. J Cell Biol. 1979 Feb;80(2):266–276. doi: 10.1083/jcb.80.2.266. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. King S. M., Otter T., Witman G. B. Purification and characterization of Chlamydomonas flagellar dyneins. Methods Enzymol. 1986;134:291–306. doi: 10.1016/0076-6879(86)34097-7. [DOI] [PubMed] [Google Scholar]
  20. Kuznetsov S. A., Gelfand V. I. Bovine brain kinesin is a microtubule-activated ATPase. Proc Natl Acad Sci U S A. 1986 Nov;83(22):8530–8534. doi: 10.1073/pnas.83.22.8530. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  22. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  23. Lee-Eiford A., Ow R. A., Gibbons I. R. Specific cleavage of dynein heavy chains by ultraviolet irradiation in the presence of ATP and vanadate. J Biol Chem. 1986 Feb 15;261(5):2337–2342. [PubMed] [Google Scholar]
  24. Luca F. C., Bloom G. S., Vallee R. B. A monoclonal antibody that cross-reacts with phosphorylated epitopes on two microtubule-associated proteins and two neurofilament polypeptides. Proc Natl Acad Sci U S A. 1986 Feb;83(4):1006–1010. doi: 10.1073/pnas.83.4.1006. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Murphy D. B., Hiebsch R. R., Wallis K. T. Identity and Origin of the ATPase activity associated with neuronal microtubules. I. The ATPase activity is associated with membrane vesicles. J Cell Biol. 1983 May;96(5):1298–1305. doi: 10.1083/jcb.96.5.1298. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Murphy D. B., Wallis K. T., Hiebsch R. R. Identity and origin of the ATPase activity associated with neuronal microtubules. II. Identification of a 50,000-dalton polypeptide with ATPase activity similar to F-1 ATPase from mitochondria. J Cell Biol. 1983 May;96(5):1306–1315. doi: 10.1083/jcb.96.5.1306. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Omoto C. K., Johnson K. A. Activation of the dynein adenosinetriphosphatase by microtubules. Biochemistry. 1986 Jan 28;25(2):419–427. doi: 10.1021/bi00350a022. [DOI] [PubMed] [Google Scholar]
  28. Pallini V., Mencarelli C., Bracci L., Contorni M., Ruggiero P., Tiezzi A., Manetti R. Cytoplasmic nucleoside-triphosphatase similar to axonemal dynein occur widely in different cell types. J Submicrosc Cytol. 1983 Jan;15(1):229–235. [PubMed] [Google Scholar]
  29. Pfister K. K., Fay R. B., Witman G. B. Purification and polypeptide composition of dynein ATPases from Chlamydomonas flagella. Cell Motil. 1982;2(6):525–547. doi: 10.1002/cm.970020604. [DOI] [PubMed] [Google Scholar]
  30. Pratt M. M., Otter T., Salmon E. D. Dynein-like Mg2+-ATPase in mitotic spindles isolated from sea urchin embryos (Strongylocentrotus droebachiensis). J Cell Biol. 1980 Sep;86(3):738–745. doi: 10.1083/jcb.86.3.738. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pratt M. M. Stable complexes of axoplasmic vesicles and microtubules: protein composition and ATPase activity. J Cell Biol. 1986 Sep;103(3):957–968. doi: 10.1083/jcb.103.3.957. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pratt M. M. The identification of a dynein ATPase in unfertilized sea urchin eggs. Dev Biol. 1980 Feb;74(2):364–378. doi: 10.1016/0012-1606(80)90438-8. [DOI] [PubMed] [Google Scholar]
  33. 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]
  34. Vale R. D., Schnapp B. J., Mitchison T., Steuer E., Reese T. S., Sheetz M. P. Different axoplasmic proteins generate movement in opposite directions along microtubules in vitro. Cell. 1985 Dec;43(3 Pt 2):623–632. doi: 10.1016/0092-8674(85)90234-x. [DOI] [PubMed] [Google Scholar]
  35. Vale R. D., Schnapp B. J., Reese T. S., Sheetz M. P. Organelle, bead, and microtubule translocations promoted by soluble factors from the squid giant axon. Cell. 1985 Mar;40(3):559–569. doi: 10.1016/0092-8674(85)90204-1. [DOI] [PubMed] [Google Scholar]
  36. Vallee R. B. A taxol-dependent procedure for the isolation of microtubules and microtubule-associated proteins (MAPs). J Cell Biol. 1982 Feb;92(2):435–442. doi: 10.1083/jcb.92.2.435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Vallee R. B., Borisy G. G. Removal of the projections from cytoplasmic microtubules in vitro by digestion with trypsin. J Biol Chem. 1977 Jan 10;252(1):377–382. [PubMed] [Google Scholar]
  38. Vallee R. B., Collins C. A. Purification of microtubules and microtubule-associated proteins from sea urchin eggs and cultured mammalian cells using taxol, and use of exogenous taxol-stabilized brain microtubules for purifying microtubule-associated proteins. Methods Enzymol. 1986;134:116–127. doi: 10.1016/0076-6879(86)34080-1. [DOI] [PubMed] [Google Scholar]
  39. Vallee R. B., Davis S. E. Low molecular weight microtubule-associated proteins are light chains of microtubule-associated protein 1 (MAP 1). Proc Natl Acad Sci U S A. 1983 Mar;80(5):1342–1346. doi: 10.1073/pnas.80.5.1342. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Vallee R. B. Reversible assembly purification of microtubules without assembly-promoting agents and further purification of tubulin, microtubule-associated proteins, and MAP fragments. Methods Enzymol. 1986;134:89–104. doi: 10.1016/0076-6879(86)34078-3. [DOI] [PubMed] [Google Scholar]
  41. Vallee R. Structure and phosphorylation of microtubule-associated protein 2 (MAP 2). Proc Natl Acad Sci U S A. 1980 Jun;77(6):3206–3210. doi: 10.1073/pnas.77.6.3206. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. White H. D., Coughlin B. A., Purich D. L. Adenosine triphosphatase activity of bovine brain microtubule protein. J Biol Chem. 1980 Jan 25;255(2):486–491. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES