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Microtubule attachment and spindle assembly checkpoint signalling at the kinetochore

Nature Reviews Molecular Cell Biology volume 14, pages 25–37 (2013)Cite this article

Subjects

Key Points

  • During mitosis, replicated sister chromatids attach to the mitotic spindle to facilitate their equal partitioning into two daughter cells.

  • Error-free chromosome segregation depends on the establishment of proper interactions between chromosomes and spindle microtubules, and on the delay of chromosome segregation, which is mediated by the activation of spindle assembly checkpoint (SAC), until all chromosomes have attached to the spindle. Both of these processes depend on the kinetochore, an assembly of proteins that connects centromeric DNA to spindle microtubules.

  • The KMN (kinetochore null protein 1 (KNL1)–missegregation 12 (MIS12)–nuclear division cycle 80 (NDC80) protein network constitutesan essential and conserved microtubule-binding activity at the kinetochore. Additionally, the KMN network is a platform for the recruitment of SAC proteins to the kinetochore, an essential step in the activation of checkpoint signalling.

  • Kinetochore–microtubule interactions are regulated by reversible protein phosphorylation, with essential contributions from kinases and phosphatases, and by protein–protein interactions. This ensures that proper spindle attachments persist until anaphase, whereas improper attachments are eliminated.

  • Phosphorylation-dependent protein–protein interactions between SAC proteins and the KMN network are important to coordinate the absence or presence of kinetochore-bound microtubules to regulate the activation or extinction of SAC signalling, respectively.

  • A complete understanding of how proper chromosome–spindle attachments are established and how their presence is relayed to the SAC to ensure error-free chromosome segregation will provide an integrated understanding of the molecular transactions at the kinetochore, together with its micromechanical properties, which function cooperatively to increase the fidelity of chromosome segregation.

Abstract

In eukaryotes, chromosome segregation during cell division is facilitated by the kinetochore, a multiprotein structure that is assembled on centromeric DNA. The kinetochore attaches chromosomes to spindle microtubules, modulates the stability of these attachments and relays the microtubule-binding status to the spindle assembly checkpoint (SAC), a cell cycle surveillance pathway that delays chromosome segregation in response to unattached kinetochores. Recent studies are shaping current thinking on how each of these kinetochore-centred processes is achieved, and how their integration ensures faithful chromosome segregation, focusing on the essential roles of kinase–phosphatase signalling and the microtubule-binding KMN protein network.

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Figure 1: Speed and fidelity in cell division.
Figure 2: Architectural organization of the KMN network.
Figure 3: Molecular mechanism of kinetochore-based SAC activation.
Figure 4: Molecular mechanisms of kinetochore–microtubule attachment.
Figure 5: Molecular mechanisms of SAC extinction at the kinetochore

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Acknowledgements

T.M.K. is grateful to the US National Institutes of Health (NIH) (GM98579). The authors apologize to their colleagues whose work could not be discussed owing to space limitations.

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  1. Cell Biology Program, Sloan-Kettering Institute, 1275 York Ave, New York, 10065, New York, USA

    Emily A. Foley

  2. Laboratory of Chemistry and Cell Biology, Rockefeller University, 1230 York Ave, New York, 10065, New York, USA

    Tarun M. Kapoor

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Glossary

Cytokinesis

The division of the cytoplasm to generate two daughter cells. It typically follows chromosome segregation.

Sister chromatids

Pairs of identical DNA sequences that are formed as a result of DNA replication. Sister chromatids are held together by cohesin complexes.

Cohesin

A multiprotein complex that tethers replicated sister chromatids. Cohesin enables sister chromatids to resist separation even when exposed to microtubule-dependent pulling forces, possibly by encircling DNA. Proteolytic cleavage of cohesin by the enzyme separase allows chromosome segregation at anaphase.

Centromeric DNA

A specialized chromosomal locus that is epigenetically defined by the presence of the histone H3 variant centromere-associated protein A (CENPA), which directs the assembly of the kinetochore.

Constitutive centromeric-associated network

(CCAN). A conserved network of proteins that is assembled on centromeric DNA. It is required for the recruitment of most kinetochore proteins.

Coiled-coil domains

Secondary structures composed of two or more -helices that entwine into a supercoil. These structures often mediate proteinprotein interactions and oligomerization.

Calponin-homology domain

A protein module of ∼110 amino acids that is found in many cytoskeletal and signal transduction proteins.

E-hook

Carboxy-terminal residues of -tubulin and -tubulin. The E-hook contains the acidic residues Glu and Asp.

Microtubule plus ends

The ends of microtubule polymers with -tubulin subunits being exposed. They are more dynamic than the microtubule minus end (in which -tubulin is exposed). In cells, microtubule nucleation occurs only at the plus end.

Cytoplasmic dynein

A multi-subunit, AAA+ ATPase, minus-end directed microtubule motor. During mitosis, it is essential for proper kinetochoremicrotubule attachments and spindle pole organization.

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Foley, E., Kapoor, T. Microtubule attachment and spindle assembly checkpoint signalling at the kinetochore. Nat Rev Mol Cell Biol 14, 25–37 (2013). https://doi.org/10.1038/nrm3494

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