Properties of the kinetochore in vitro. I. Microtubule nucleation and tubulin binding

doi:10.1083/jcb.101.3.755

This Article

	Full Text (PDF, 1377K)
	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 Mitchison, T. J.
	Articles by Kirschner, M. W.
	Search for Related Content

PubMed

	PubMed Citation
	Articles by Mitchison, T. J.
	Articles by Kirschner, M. W.


Social Bookmarking

	What's this?

ARTICLES

Properties of the kinetochore in vitro. I. Microtubule nucleation and tubulin binding

TJ Mitchison and MW Kirschner

We have isolated chromosomes from Chinese hamster ovary cells arrested in mitosis with vinblastine and examined the interactions of their kinetochores with purified tubulin in vitro. The kinetochores nucleate microtubule (MT) growth with complex kinetics. After an initial lag phase, MTs are continuously nucleated with both plus and minus ends distally localized. This mixed polarity seems inconsistent with the formation of an ordered, homopolar kinetochore fiber in vivo. As isolated from vinblastine-arrested cells, kinetochores contain no bound tubulin. The kinetochores of chromosomes isolated from colcemid- arrested cells or of chromosomes incubated with tubulin in vitro are brightly stained after anti-tubulin immunofluorescence. This bound tubulin is probably not in the form of MTs. It is localized to the corona region by immunoelectron microscopy, where it may play a role in MT nucleation in vitro.
Add to CiteULike CiteULike Add to Complore Complore Add to Connotea Connotea Add to Del.icio.us Del.icio.us Add to Digg Digg Add to Reddit Reddit Add to Technorati Technorati What's this?

This article has been cited by other articles:

Dhonukshe, P., Vischer, N., Gadella, T. W. J. Jr (2006). Contribution of microtubule growth polarity and flux to spindle assembly and functioning in plant cells.. J. Cell Sci. 119: 3193-3205 [Abstract] [Full Text]
Powell, K. (2006). Chromosome snatching by lateral microtubules. JCB 172: 788-788 [Full Text]
Maiato, H., DeLuca, J., Salmon, E. D., Earnshaw, W. C. (2004). The dynamic kinetochore-microtubule interface. J. Cell Sci. 117: 5461-5477 [Abstract] [Full Text]
Hunt, A. J., McIntosh, J. R. (1998). The Dynamic Behavior of Individual Microtubules Associated with Chromosomes In Vitro. Mol. Biol. Cell 9: 2857-2871 [Abstract] [Full Text]
Desai, A., Deacon, H. W., Walczak, C. E., Mitchison, T. J. (1997). A method that allows the assembly of kinetochore components onto chromosomes condensed in clarified Xenopus egg�extracts. Proc. Natl. Acad. Sci. USA 94: 12378-12383 [Abstract] [Full Text]
Yao, X., Anderson, K. L., Cleveland, D. W. (1997). The Microtubule-dependent Motor Centromere-associated Protein E (CENP-E) Is an Integral Component of Kinetochore Corona Fibers That Link Centromeres to Spindle Microtubules. JCB 139: 435-447 [Abstract] [Full Text]
Minin, A. (1997). Dispersal of Golgi apparatus in nocodazole-treated fibroblasts is a kinesin-driven process. J. Cell Sci. 110: 2495-2505 [Abstract]
Pluta, A. F., Earnshaw, W. C. (1996). Specific Interaction between Human Kinetochore Protein CENP-C and a Nucleolar Transcriptional Regulator. J. Biol. Chem. 271: 18767-18774 [Abstract] [Full Text]
Brown, K., Wood, K., Cleveland, D. (1996). The kinesin-like protein CENP-E is kinetochore-associated throughout poleward chromosome segregation during anaphase-A. J. Cell Sci. 109: 961-969 [Abstract]
Williams, B., Goldberg, M. (1994). Determinants of Drosophila zw10 protein localization and function. J. Cell Sci. 107: 785-798 [Abstract]
Kuo, S., Sheetz, M. (1993). Force of single kinesin molecules measured with optical tweezers. Science 260: 232-234 [Abstract]
Rodionov, V., Gelfand, V., Borisy, G. (1993). Kinesin-like molecules involved in spindle formation. J. Cell Sci. 106: 1179-1188 [Abstract]
Nicklas, R., Krawitz, L., Ward, S. (1993). Odd chromosome movement and inaccurate chromosome distribution in mitosis and meiosis after treatment with protein kinase inhibitors. J. Cell Sci. 104: 961-973 [Abstract]
Rattner, J. B., Wang, T. (1992). Kinetochore formation and behaviour following premature chromosome condensation. J. Cell Sci. 103: 1039-1045 [Abstract]
Earnshaw, W.C., Bernat, R.L., Cooke, C.A., Rothfield, N.F. (1991). Role of the Centromere/Kinetochore in Cell Cycle Control. Cold Spring Harb Symp Quant Biol 56: 675-685 [Abstract]
Hyman, A.A., Mitchison, T.J. (1991). Regulation of the Direction of Chromosome Movement. Cold Spring Harb Symp Quant Biol 56: 745-750 [Abstract]
McIntosh, J., Koonce, M. (1989). Mitosis. Science 246: 622-628 [Abstract]