A Biosensor for the Mitotic Kinase MPS1 Reveals Spatiotemporal Activity Dynamics and Regulation

Kuijt, Timo E. F.; Lambers, Maaike L.A.; Weterings, Sonja; Ponsioen, Bas; Bolhaqueiro, Ana C.F.; Staijen, Debbie H.M.; Kops, Geert J.P.L.

doi:10.1016/j.cub.2020.07.062

Cited by 28 publications

(26 citation statements)

References 70 publications

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“…A crucial factor likely explaining why RZZS filaments form only at kinetochores is that RZZS polymerization appears to be kinetically controlled, with MPS1 phosphorylation acting as catalyst to remove a steric blockade to oligomerization involving the ROD N‐terminal region. As kinetochores enrich MPS1 during mitosis, and MPS1 activity is highest at these structures, albeit not limited to them (Kuijt et al , 2020), polymerization may become naturally spatially limited to kinetochores. The release of ROD from an auto‐inhibited state upon MPS1 phosphorylation, postulated here, should be considered alongside the proposed auto‐inhibition of Spindly, shown to involve an intramolecular interaction of its coiled coils (Sacristan et al , 2018; preprint: d’Amico et al , 2022).…”

Section: Discussionmentioning

confidence: 99%

Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores

et al. 2022

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In metazoans, a ≈1 megadalton (MDa) multiprotein complex comprising the dynein–dynactin adaptor Spindly and the ROD–Zwilch–ZW10 (RZZ) complex is the building block of a fibrous biopolymer, the kinetochore fibrous corona. The corona assembles on mitotic kinetochores to promote microtubule capture and spindle assembly checkpoint (SAC) signaling. We report here a high‐resolution cryo‐EM structure that captures the essential features of the RZZ complex, including a farnesyl‐binding site required for Spindly binding. Using a highly predictive in vitro assay, we demonstrate that the SAC kinase MPS1 is necessary and sufficient for corona assembly at supercritical concentrations of the RZZ–Spindly (RZZS) complex, and describe the molecular mechanism of phosphorylation‐dependent filament nucleation. We identify several structural requirements for RZZS polymerization in rings and sheets. Finally, we identify determinants of kinetochore localization and corona assembly of Spindly. Our results describe a framework for the long‐sought‐for molecular basis of corona assembly on metazoan kinetochores.

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Section: Discussionmentioning

confidence: 99%

Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores

et al. 2022

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“…A second crucial factor likely explaining why RZZS filaments form only at kinetochores is that RZZS polymerization appears to be kinetically controlled, with MPS1 phosphorylation acting as catalyst to remove a steric blockade to oligomerization involving the ROD N-terminal region. As kinetochores enrich MPS1 during mitosis, and MPS1 activity is highest at these structures, albeit not limited to them (Kuijt et al, 2020), polymerization may become naturally spatially limited to kinetochores.…”

Section: Discussionmentioning

confidence: 99%

Structure of the RZZ complex and molecular basis of Spindly-driven corona assembly at human kinetochores

Raisch

Ciossani

d'Amico

et al. 2021

Preprint

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In metazoans, a ≍1 megadalton (MDa) super-complex comprising the Dynein-Dynactin adaptor Spindly and the ROD-Zwilch-ZW10 (RZZ) complex is the building block of a fibrous biopolymer, the kinetochore fibrous corona. The corona assembles on mitotic kinetochores to promote microtubule capture and spindle assembly checkpoint (SAC) signaling. We report here a high-resolution cryo-EM structure that captures the essential features of the RZZ complex, including a farnesyl binding site required for Spindly binding. Using a highly predictive in vitro assay, we demonstrate that the SAC kinase MPS1 is necessary and sufficient for corona assembly at supercritical concentrations of the RZZ-Spindly (RZZS) complex, and describe the molecular mechanism of phosphorylation-dependent filament nucleation. We identify several structural requirements for RZZS polymerization in rings and sheets. Finally, we identify determinants of kinetochore localization and corona assembly of Spindly. Our results describe a framework for the long-sought-for molecular basis of corona assembly on metazoan kinetochores.

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“…Consistent with this interpretation, inhibition of Mps1 does not alter the phosphorylation of many Aurora B substrates or Aurora B localization (Hewitt et al, 2010;Maciejowski et al, 2010;Maure et al, 2007;Santaguida et al, 2010;Tighe et al, 2008). Mps1 localizes to kinetochores by binding to the Ndc80 protein, and its activity is highest on kinetochores that have not made proper attachments (Hiruma et al, 2015;Ji et al, 2015;Kemmler et al, 2009;Kuijt et al, 2020). However, its role in regulating kinetochore-microtubule attachments and error correction has not been fully elucidated.…”

Section: Introductionmentioning

confidence: 90%

Kinetochore-bound Mps1 regulates kinetochore–microtubule attachments via Ndc80 phosphorylation

Sarangapani

Koch

Nelson

et al. 2021

Journal of Cell Biology

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Dividing cells detect and correct erroneous kinetochore–microtubule attachments during mitosis, thereby avoiding chromosome missegregation. The Aurora B kinase phosphorylates microtubule-binding elements specifically at incorrectly attached kinetochores, promoting their release and providing another chance for proper attachments to form. However, growing evidence suggests that the Mps1 kinase is also required for error correction. Here we directly examine how Mps1 activity affects kinetochore–microtubule attachments using a reconstitution-based approach that allows us to separate its effects from Aurora B activity. When endogenous Mps1 that copurifies with kinetochores is activated in vitro, it weakens their attachments to microtubules via phosphorylation of Ndc80, a major microtubule-binding protein. This phosphorylation contributes to error correction because phospho-deficient Ndc80 mutants exhibit genetic interactions and segregation defects when combined with mutants in other error correction pathways. In addition, Mps1 phosphorylation of Ndc80 is stimulated on kinetochores lacking tension. These data suggest that Mps1 provides an additional mechanism for correcting erroneous kinetochore–microtubule attachments, complementing the well-known activity of Aurora B.

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A Biosensor for the Mitotic Kinase MPS1 Reveals Spatiotemporal Activity Dynamics and Regulation

Cited by 28 publications

References 70 publications

Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores

Structure of the RZZ complex and molecular basis of Spindly‐driven corona assembly at human kinetochores

Structure of the RZZ complex and molecular basis of Spindly-driven corona assembly at human kinetochores

Kinetochore-bound Mps1 regulates kinetochore–microtubule attachments via Ndc80 phosphorylation

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