Access

You are not currently logged in.

Access your personal account or get JSTOR access through your library or other institution:

login

Log in to your personal account or through your institution.

If You Use a Screen Reader

This content is available through Read Online (Free) program, which relies on page scans. Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.

A Method that Allows the Assembly of Kinetochore Components onto Chromosomes Condensed in Clarified Xenopus Egg Extracts

Arshad Desai, Heather W. Deacon, Claire E. Walczak and Timothy J. Mitchison
Proceedings of the National Academy of Sciences of the United States of America
Vol. 94, No. 23 (Nov. 11, 1997), pp. 12378-12383
Stable URL: http://www.jstor.org/stable/43915
Page Count: 6
  • Read Online (Free)
  • Subscribe ($19.50)
  • Cite this Item
Since scans are not currently available to screen readers, please contact JSTOR User Support for access. We'll provide a PDF copy for your screen reader.
A Method that Allows the Assembly of Kinetochore Components onto Chromosomes Condensed in Clarified Xenopus Egg Extracts
Preview not available

Abstract

Kinetochores are complex macromolecular structures that link mitotic chromosomes to spindle microtubules. Although a small number of kinetochore components have been identified, including the kinesins CENP-E and XKCM1 as well as cytoplasmic dynein, neither how these and other proteins are organized to produce a kinetochore nor their exact functions within this structure are understood. For this reason, we have developed an assay that allows kinetochore components to assemble onto discrete foci on in vitro-condensed chromosomes. The source of the kinetochore components is a clarified cell extract from Xenopus eggs that can be fractionated or immunodepleted of individual proteins. Kinetochore assembly in these clarified extracts requires preincubating the substrate sperm nuclei in an extract under low ATP conditions. Immunodepletion of XKCM1 from the extracts prevents the localization of kinetochore-associated XKCM1 without affecting the targeting of CENP-E and cytoplasmic dynein or the binding of monomeric tubulin to the kinetochore. Extension of this assay for the analysis of other components should help to dissect the protein-protein interactions involved in kinetochore assembly and function.

Page Thumbnails

  • Thumbnail: Page 
12378
    12378
  • Thumbnail: Page 
12379
    12379
  • Thumbnail: Page 
12380
    12380
  • Thumbnail: Page 
12381
    12381
  • Thumbnail: Page 
12382
    12382
  • Thumbnail: Page 
12383
    12383