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J Cell Biol. 2013 Sep 2;202(5):735-46. doi: 10.1083/jcb.201304197. Epub 2013 Aug 26.

Spindle assembly checkpoint proteins are positioned close to core microtubule attachment sites at kinetochores.

Author information

1
Department of Biology, The University of North Carolina-Chapel Hill, Chapel Hill, NC 27599, USA. dileep@email.unc.edu

Abstract

Spindle assembly checkpoint proteins have been thought to reside in the peripheral corona region of the kinetochore, distal to microtubule attachment sites at the outer plate. However, recent biochemical evidence indicates that checkpoint proteins are closely linked to the core kinetochore microtubule attachment site comprised of the Knl1-Mis12-Ndc80 (KMN) complexes/KMN network. In this paper, we show that the Knl1-Zwint1 complex is required to recruit the Rod-Zwilch-Zw10 (RZZ) and Mad1-Mad2 complexes to the outer kinetochore. Consistent with this, nanometer-scale mapping indicates that RZZ, Mad1-Mad2, and the C terminus of the dynein recruitment factor Spindly are closely juxtaposed with the KMN network in metaphase cells when their dissociation is blocked and the checkpoint is active. In contrast, the N terminus of Spindly is ∼75 nm outside the calponin homology domain of the Ndc80 complex. These results reveal how checkpoint proteins are integrated within the substructure of the kinetochore and will aid in understanding the coordination of microtubule attachment and checkpoint signaling during chromosome segregation.

PMID:
23979716
PMCID:
PMC3760617
DOI:
10.1083/jcb.201304197
[Indexed for MEDLINE]
Free PMC Article

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