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PMID: 9786942 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't Research Support, U.S. Gov't, P.H.S.

Spindle checkpoint protein Xmad1 recruits Xmad2 to unattached kinetochores.

The Journal of cell biology ·Vol. 143 ·No. 2 ·1998-10-19 ·Pages 283-95

Chen RH, Shevchenko A, Mann M, Murray AW

Abstract

The spindle checkpoint prevents the metaphase to anaphase transition in cells containing defects in the mitotic spindle or in chromosome attachment to the spindle. When the checkpoint protein Xmad2 is depleted from Xenopus egg extracts, adding Xmad2 to its endogenous concentration fails to restore the checkpoint, suggesting that other checkpoint component(s) were depleted from the extract through their association with Xmad2. Mass spectrometry provided peptide sequences from an 85-kD protein that coimmunoprecipitates with Xmad2 from egg extracts. This information was used to clone XMAD1, which encodes a homologue of the budding yeast (Saccharomyces cerevisiae) checkpoint protein Mad1. Xmad1 is essential for establishing and maintaining the spindle checkpoint in egg extracts. Like Xmad2, Xmad1 localizes to the nuclear envelope and the nucleus during interphase, and to those kinetochores that are not bound to spindle microtubules during mitosis. Adding an anti-Xmad1 antibody to egg extracts inactivates the checkpoint and prevents Xmad2 from localizing to unbound kinetochores. In the presence of excess Xmad2, neither chromosomes nor Xmad1 are required to activate the spindle checkpoint, suggesting that the physiological role of Xmad1 is to recruit Xmad2 to kinetochores that have not bound microtubules.

MeSH Terms
Animals DNA-Binding Proteins/genetics,metabolism HeLa Cells Humans Kinetochores/physiology Metaphase/physiology Microscopy, Fluorescence Microtubules/physiology Mitosis/physiology Molecular Sequence Data Sequence Homology, Amino Acid Signal Transduction/physiology Smad Proteins Smad2 Protein Spindle Apparatus/physiology Trans-Activators/genetics,metabolism Xenopus Proteins Xenopus laevis
Chemicals
DNA-Binding Proteins MXD1 protein, Xenopus SMAD2 protein, human Smad Proteins Smad2 Protein Smad2 protein, Xenopus Trans-Activators Xenopus Proteins
Authors & Affiliations
4 authors, click to expand affiliations / ORCID
Chen R H
Section of Biochemistry, Molecular and Cell Biology, Cornell University, Ithaca, New York 14853, USA. rc70@cornell.edu
Shevchenko A
Mann M
Murray A W
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Article Info
Journal
The Journal of cell biology
Abbr.
J Cell Biol
ISSN
0021-9525
DOI
10.1083/jcb.143.2.283
Email
rc70@cornell.edu
Published
1998-10-19
Pages
283-95
Language
English
Region
United States
NLM ID
0375356
PMCID
PMC2132829
Subset
IM
Databases
GENBANK
AF126481
External Links
PubMed source DOI: 10.1083/jcb.143.2.283 PMC Full Text View journal details
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