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PMID: 11707408 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

Mad2 binding to Mad1 and Cdc20, rather than oligomerization, is required for the spindle checkpoint.

The EMBO journal ·Vol. 20 ·No. 22 ·2001-11-15 ·Pages 6371-82

Sironi L, Melixetian M, Faretta M, Prosperini E, Helin K, Musacchio A

Abstract

Mad2 is a key component of the spindle checkpoint, a device that controls the fidelity of chromosome segregation in mitosis. The ability of Mad2 to form oligomers in vitro has been correlated with its ability to block the cell cycle upon injection into Xenopus embryos. Here we show that Mad2 forms incompatible complexes with Mad1 and Cdc20, neither of which requires Mad2 oligomerization. A monomeric point mutant of Mad2 can sustain a cell cycle arrest of comparable strength to that of the wild-type protein. We show that the interaction of Mad2 with Mad1 is crucial for the localization of Mad2 to kinetochores, where Mad2 interacts with Cdc20. We propose a model that features the kinetochore as a 'folding factory' for the formation of a Mad2-Cdc20 complex endowed with inhibitory activity on the anaphase promoting complex.

MeSH Terms
Anaphase Animals Antibodies, Monoclonal/metabolism Binding Sites Calcium-Binding Proteins/metabolism Carrier Proteins Cdc20 Proteins Cell Cycle Cell Cycle Proteins/metabolism Chromatography, Gel DNA, Complementary/metabolism Electrophoresis, Polyacrylamide Gel Escherichia coli/metabolism Fungal Proteins/metabolism HeLa Cells Humans Kinetochores/metabolism Mad2 Proteins Mice Microscopy, Fluorescence Mitosis Models, Biological Mutagenesis, Site-Directed Nuclear Proteins Peptides/chemistry Phosphoproteins/metabolism Plasmids/metabolism Point Mutation Polymerase Chain Reaction Precipitin Tests Protein Binding Recombinant Proteins/metabolism Repressor Proteins/metabolism Saccharomyces cerevisiae Proteins Sodium Chloride/pharmacology Transfection Urea/pharmacology
Chemicals
Antibodies, Monoclonal CDC20 protein, S cerevisiae Calcium-Binding Proteins Carrier Proteins Cdc20 Proteins Cell Cycle Proteins DNA, Complementary Fungal Proteins MAD1 protein, S cerevisiae MAD1L1 protein, human MAD2 protein, S cerevisiae Mad1l1 protein, mouse Mad2 Proteins Mad2l1 protein, mouse Mad2l2 protein, mouse Nuclear Proteins Peptides Phosphoproteins Recombinant Proteins Repressor Proteins Saccharomyces cerevisiae Proteins Sodium Chloride Urea
Authors & Affiliations
6 authors, click to expand affiliations / ORCID
Sironi L
Department of Experimental Oncology, European Institute of Oncology, Via Ripamonti 435, 20141 Milan, Italy.
Melixetian M
Faretta M
Prosperini E
Helin K
Musacchio A
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37 references, click to expand
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Article Info
Journal
The EMBO journal
Abbr.
EMBO J
ISSN
0261-4189
DOI
10.1093/emboj/20.22.6371
Published
2001-11-15
Pages
6371-82
Language
English
Region
England
NLM ID
8208664
PMCID
PMC125308
Subset
IM
External Links
PubMed source DOI: 10.1093/emboj/20.22.6371 PMC Full Text View journal details
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