Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP 1C)

doi:10.1083/jcb.107.3.1001

Quantitative Colocalization Analysis Software

This Article

	Full Text (PDF, 996K)
	Alert me when this article is cited
	Citation Map

Services

	Email this article
	Similar articles in this journal
	Similar articles in PubMed
	Alert me to new content in the JCB
	Download to citation manager

Citing Articles

	Citing Articles via HighWire
	Citing Articles via CrossRef
	Citing Articles via Google Scholar

Google Scholar

	Articles by Shpetner, H. S.
	Articles by Vallee, R. B.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Shpetner, H. S.
	Articles by Vallee, R. B.


Social Bookmarking

	What's this?

ARTICLES

Characterization of the microtubule-activated ATPase of brain cytoplasmic dynein (MAP 1C)

HS Shpetner, BM Paschal and RB Vallee
Cell Biology Group, Worcester Foundation for Experimental Biology, Shrewsbury, Massachusetts 01545.

We recently found that the brain cytosolic microtubule-associated protein 1C (MAP 1C) is a microtubule-activated ATPase, capable of translocating microtubules in vitro in the direction corresponding to retrograde transport. (Paschal, B. M., H. S. Shpetner, and R. B. Vallee. 1987b. J. Cell Biol. 105:1273-1282; Paschal, B. M., and R. B. Vallee. 1987. Nature [Lond.]. 330:181-183.). Biochemical analysis of this protein (op. cit.) as well as scanning transmission electron microscopy revealed that MAP 1C is a brain cytoplasmic form of the ciliary and flagellar ATPase dynein (Vallee, R. B., J. S. Wall, B. M. Paschal, and H. S. Shpetner. 1988. Nature [Lond.]. 332:561-563). We have now characterized the ATPase activity of the brain enzyme in detail. We found that microtubule activation required polymeric tubulin and saturated with increasing tubulin concentration. The maximum activity at saturating tubulin (Vmax) varied from 186 to 239 nmol/min per mg. At low ionic strength, the Km for microtubules was 0.16 mg/ml tubulin, substantially lower than that previously reported for axonemal dynein. The microtubule-stimulated activity was extremely sensitive to changes in ionic strength and sulfhydryl oxidation state, both of which primarily affected the microtubule concentrations required for half- maximal activation. In a number of respects the brain dynein was enzymatically similar to both axonemal and egg dyneins. Thus, the ATPase required divalent cations, calcium stimulating activity less effectively than magnesium. The MgATPase was inhibited by metavandate (Ki = 5-10 microM for the microtubule-stimulated activity), 1 mM NEM, and 1 mM EHNA. In contrast to other dyneins, the brain enzyme hydrolyzed CTP, TTP, and GTP at higher rates than ATP. Thus, the enzymological properties of the brain cytoplasmic dynein are clearly related to those of other dyneins, though the brain enzyme is unique in its substrate specificity and in its high sensitivity to stimulation by microtubules.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Facebook Add to Reddit Reddit Add to Technorati Technorati Twitter What's this?

This article has been cited by other articles:

Willis, D. E., van Niekerk, E. A., Sasaki, Y., Mesngon, M., Merianda, T. T., Williams, G. G., Kendall, M., Smith, D. S., Bassell, G. J., Twiss, J. L. (2007). Extracellular stimuli specifically regulate localized levels of individual neuronal mRNAs. JCB 178: 965-980 [Abstract] [Full Text]
Kelkar, S., De, B. P., Gao, G., Wilson, J. M., Crystal, R. G., Leopold, P. L. (2006). A Common Mechanism for Cytoplasmic Dynein-Dependent Microtubule Binding Shared among Adeno-Associated Virus and Adenovirus Serotypes.. J. Virol. 80: 7781-7785 [Abstract] [Full Text]
Hook, P., Mikami, A., Shafer, B., Chait, B. T., Rosenfeld, S. S., Vallee, R. B. (2005). Long Range Allosteric Control of Cytoplasmic Dynein ATPase Activity by the Stalk and C-terminal Domains. J. Biol. Chem. 280: 33045-33054 [Abstract] [Full Text]
Kelkar, S. A., Pfister, K. K., Crystal, R. G., Leopold, P. L. (2004). Cytoplasmic Dynein Mediates Adenovirus Binding to Microtubules. J. Virol. 78: 10122-10132 [Abstract] [Full Text]
Nishiura, M., Kon, T., Shiroguchi, K., Ohkura, R., Shima, T., Toyoshima, Y. Y., Sutoh, K. (2004). A Single-headed Recombinant Fragment of Dictyostelium Cytoplasmic Dynein Can Drive the Robust Sliding of Microtubules. J. Biol. Chem. 279: 22799-22802 [Abstract] [Full Text]
Reck-Peterson, S. L., Vale, R. D. (2004). Molecular dissection of the roles of nucleotide binding and hydrolysis in dynein's AAA domains in Saccharomyces cerevisiae. Proc. Natl. Acad. Sci. USA 101: 1491-1495 [Abstract] [Full Text]
Bailey, C. J., Crystal, R. G., Leopold, P. L. (2003). Association of Adenovirus with the Microtubule Organizing Center. J. Virol. 77: 13275-13287 [Abstract] [Full Text]
Roobol, A., Sahyoun, Z. P., Carden, M. J. (1999). Selected Subunits of the Cytosolic Chaperonin Associate with Microtubules Assembled in Vitro. J. Biol. Chem. 274: 2408-2415 [Abstract] [Full Text]
Palazzo, R., Vaisberg, E., Weiss, D., Kuznetsov, S., Steffen, W (1999). Dynein is required for spindle assembly in cytoplasmic extracts of Spisula solidissima oocytes. J. Cell Sci. 112: 1291-1302 [Abstract]
Fullerton, A. T., Bau, M.-Y., Conrad, P. A., Bloom, G. S. (1998). In Vitro Reconstitution of Microtubule Plus End-directed, GTPgamma S-sensitive Motility ofGolgi Membranes. Mol. Biol. Cell 9: 2699-2714 [Abstract] [Full Text]
Lochner, J. E., Kingma, M., Kuhn, S., Meliza, C. D., Cutler, B., Scalettar, B. A. (1998). Real-Time Imaging of the Axonal Transport of Granules Containing a Tissue Plasminogen Activator/Green Fluorescent Protein Hybrid. Mol. Biol. Cell 9: 2463-2476 [Abstract] [Full Text]
Criswell, P., Ostrowski, L., Asai, D. (1996). A novel cytoplasmic dynein heavy chain: expression of DHC1b in mammalian ciliated epithelial cells. J. Cell Sci. 109: 1891-1898 [Abstract]
Ferro, K. L., Collins, C. A. (1995). Microtubule-independent Phospholipid Stimulation of Cytoplasmic Dynein ATPase Activity. J. Biol. Chem. 270: 4492-4496 [Abstract] [Full Text]
Hughes, S., Vaughan, K., Herskovits, J., Vallee, R. (1995). Molecular analysis of a cytoplasmic dynein light intermediate chain reveals homology to a family of ATPases. J. Cell Sci. 108: 17-24 [Abstract]
Hays, T., Porter, M., McGrail, M, Grissom, P, Gosch, P, Fuller, M., McIntosh, J. (1994). A cytoplasmic dynein motor in Drosophila: identification and localization during embryogenesis. J. Cell Sci. 107: 1557-1569 [Abstract]
Asai, D., Beckwith, S., Kandl, K., Keating, H., Tjandra, H, Forney, J. (1994). The dynein genes of Paramecium tetraurelia. Sequences adjacent to the catalytic P-loop identify cytoplasmic and axonemal heavy chain isoforms. J. Cell Sci. 107: 839-847 [Abstract]
Wilkerson, C., King, S., Witman, G. (1994). Molecular analysis of the gamma heavy chain of Chlamydomonas flagellar outer-arm dynein. J. Cell Sci. 107: 497-506 [Abstract]