The right place at the right time: Aurora B kinase localization to centromeres and kinetochores

Abstract The fidelity of chromosome segregation during mitosis is intimately linked to the function of kinetochores, which are large protein complexes assembled at sites of centromeric heterochromatin on mitotic chromosomes. These key “orchestrators” of mitosis physically connect chromosomes to spindle microtubules and transduce forces through these connections to congress chromosomes and silence the spindle assembly checkpoint. Kinetochore-microtubule attachments are highly regulated to ensure that incorrect attachments are not prematurely stabilized, but instead released and corrected. The kinase activity of the centromeric protein Aurora B is required for kinetochore-microtubule destabilization during mitosis, but how the kinase acts on outer kinetochore substrates to selectively destabilize immature and erroneous attachments remains debated. Here, we review recent literature that sheds light on how Aurora B kinase is recruited to both centromeres and kinetochores and discuss possible mechanisms for how kinase interactions with substrates at distinct regions of mitotic chromosomes are regulated.

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Kinetochores respond to subtle changes in the stability of microtubule attachments

10.1101/2021.02.19.432040 ◽

2021 ◽

Author(s):

Jessica D. Warren ◽

Sarah Y. Valles ◽

Duane A. Compton

Keyword(s):

Chromosome Segregation ◽

Spindle Assembly Checkpoint ◽

Protein Localization ◽

Aurora B ◽

Aurora B Kinase ◽

Regulatory Enzymes ◽

Summary Statement ◽

Spindle Microtubules ◽

The Stability ◽

Induced Changes

AbstractProper attachment of spindle microtubules to kinetochores is necessary to satisfy the spindle assembly checkpoint and ensure faithful chromosome segregation. Microtubules detach from kinetochores to correct improperly oriented attachments, and overall kinetochore-microtubule (k-MT) attachment stability is determined in response to regulatory enzymes and the activities of kinetochore-associated microtubule stabilizing and destabilizing proteins. However, it is unknown whether regulatory enzyme activity or kinetochore-associated protein localization respond to subtle changes in k-MT attachment stability. To test for this feedback response, we monitored Aurora B kinase activity and the localization of select kinetochore proteins in metaphase cells following treatments that subtly stabilize or destabilize k-MT attachments using low dose Taxol or UMK57 (an MCAK agonist), respectively. Increasing k-MT stability induced changes in the abundance of some kinetochore proteins. In contrast, reducing k-MT stability induced both increases in Aurora B kinase signaling and changes in the abundance of some kinetochore proteins. Thus, kinetochores dynamically respond to changes in the stability of their attached microtubules. This feedback control contributes to tuning k-MT attachment stability required for efficient error correction to facilitate faithful chromosome segregation.Summary StatementLive cell imaging demonstrates that kinetochore signaling responds to feedback from attached microtubules to tune their stability to ensure faithful chromosome segregation during cell division.

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Centromere-localized Aurora B kinase is required for the fidelity of chromosome segregation

The Journal of Cell Biology ◽

10.1083/jcb.201907092 ◽

2019 ◽

Vol 219 (2) ◽

Cited By ~ 9

Author(s):

Cai Liang ◽

Zhenlei Zhang ◽

Qinfu Chen ◽

Haiyan Yan ◽

Miao Zhang ◽

...

Keyword(s):

Chromosome Segregation ◽

Essential Role ◽

Spindle Assembly Checkpoint ◽

Histone H3 ◽

Spindle Assembly ◽

Aurora B ◽

Aurora B Kinase ◽

Chromosome Missegregation ◽

Proper Timing ◽

Segregation Of Chromosomes

Aurora B kinase plays an essential role in chromosome bi-orientation, which is a prerequisite for equal segregation of chromosomes during mitosis. However, it remains largely unclear whether centromere-localized Aurora B is required for faithful chromosome segregation. Here we show that histone H3 Thr-3 phosphorylation (H3pT3) and H2A Thr-120 phosphorylation (H2ApT120) can independently recruit Aurora B. Disrupting H3pT3-mediated localization of Aurora B at the inner centromere impedes the decline in H2ApT120 during metaphase and causes H2ApT120-dependent accumulation of Aurora B at the kinetochore-proximal centromere. Consequently, silencing of the spindle assembly checkpoint (SAC) is delayed, whereas the fidelity of chromosome segregation is negligibly affected. Further eliminating an H2ApT120-dependent pool of Aurora B restores proper timing for SAC silencing but increases chromosome missegregation. Our data indicate that H2ApT120-mediated localization of Aurora B compensates for the loss of an H3pT3-dependent pool of Aurora B to correct improper kinetochore–microtubule attachments. This study provides important insights into how centromeric Aurora B regulates SAC and kinetochore attachment to microtubules to ensure error-free chromosome segregation.

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Interplay between mitotic kinesins and the Aurora kinase–PP1 (protein phosphatase 1) axis

Biochemical Society Transactions ◽

10.1042/bst20130191 ◽

2013 ◽

Vol 41 (6) ◽

pp. 1761-1765 ◽

Cited By ~ 11

Author(s):

John C. Meadows

Keyword(s):

Protein Phosphatase ◽

Spindle Assembly Checkpoint ◽

Spindle Checkpoint ◽

Aurora Kinase ◽

Protein Phosphatase 1 ◽

Aurora B ◽

Spatial Gradient ◽

Cell Cycle Regulators ◽

Aurora B Kinase ◽

Surveillance Mechanism

Correct transmission of genetic information from mother to daughter cells is necessary for development and survival. Accurate segregation is achieved by bipolar attachment of sister kinetochores in each chromatid pair to spindle microtubules emanating from opposite spindle poles, a process known as chromosome bi-orientation. Achieving this requires dynamic interplay between kinetochore proteins, kinesin motor proteins and cell cycle regulators. Chromosome bi-orientation is monitored by a surveillance mechanism known as the SAC (spindle assembly checkpoint). The Aurora B kinase, which is bound to the inner centromere during early mitosis, plays a central role in both chromosome bi-orientation and the spindle checkpoint. The application of tension across centromeres establishes a spatial gradient of high phosphorylation activity at the inner centromere and low phosphorylation activity at the outer kinetochore. This gradient is further refined by the association of PP1 (protein phosphatase 1) to the outer kinetochore, which stabilizes kinetochore–microtubule interactions and silences the spindle checkpoint by dephosphorylating Aurora B kinase targets when chromosome bi-orientation is achieved. In the present review, I discuss emerging evidence that bidirectional cross-talk between mitotic kinesins and the Aurora kinase–PP1 axis is crucial for co-ordinating chromosome bi-orientation and spindle checkpoint signalling in eukaryotes.

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Enrichment of Aurora B kinase at the inner kinetochore controls outer kinetochore assembly

The Journal of Cell Biology ◽

10.1083/jcb.201901004 ◽

2019 ◽

Vol 218 (10) ◽

pp. 3237-3257 ◽

Cited By ~ 10

Author(s):

Mary Kate Bonner ◽

Julian Haase ◽

Jason Swinderman ◽

Hyunmi Halas ◽

Lisa M. Miller Jenkins ◽

...

Keyword(s):

Xenopus Laevis ◽

Central Region ◽

Spindle Assembly Checkpoint ◽

Spindle Assembly ◽

Aurora B ◽

Aurora B Kinase ◽

Kinetochore Assembly ◽

Egg Extracts ◽

Chromosome Attachment

Outer kinetochore assembly enables chromosome attachment to microtubules and spindle assembly checkpoint (SAC) signaling in mitosis. Aurora B kinase controls kinetochore assembly by phosphorylating the Mis12 complex (Mis12C) subunit Dsn1. Current models propose Dsn1 phosphorylation relieves autoinhibition, allowing Mis12C binding to inner kinetochore component CENP-C. Using Xenopus laevis egg extracts and biochemical reconstitution, we found that autoinhibition of the Mis12C by Dsn1 impedes its phosphorylation by Aurora B. Our data indicate that the INCENP central region increases Dsn1 phosphorylation by enriching Aurora B at inner kinetochores, close to CENP-C. Furthermore, centromere-bound CENP-C does not exchange in mitosis, and CENP-C binding to the Mis12C dramatically increases Dsn1 phosphorylation by Aurora B. We propose that the coincidence of Aurora B and CENP-C at inner kinetochores ensures the fidelity of kinetochore assembly. We also found that the central region is required for the SAC beyond its role in kinetochore assembly, suggesting that kinetochore enrichment of Aurora B promotes the phosphorylation of other kinetochore substrates.

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Phosphorylation of CENP-C by Aurora B facilitates kinetochore attachment error correction in mitosis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1710506114 ◽

2017 ◽

Vol 114 (50) ◽

pp. E10667-E10676 ◽

Cited By ~ 11

Author(s):

Xing Zhou ◽

Fan Zheng ◽

Chengliang Wang ◽

Minhao Wu ◽

Xiaozhen Zhang ◽

...

Keyword(s):

Error Correction ◽

Chromosome Segregation ◽

Spindle Assembly Checkpoint ◽

Mitotic Chromosome ◽

Regulatory Mechanism ◽

Dynamic Interaction ◽

Aurora B ◽

Kinetochore Assembly ◽

Important Pathway ◽

Spindle Microtubules

Kinetochores are superprotein complexes that orchestrate chromosome segregation via a dynamic interaction with spindle microtubules. A physical connection between CENP-C and the Mis12–Ndc80–Knl1 (KMN) protein network is an important pathway that is used to assemble kinetochores on CENP-A nucleosomes. Multiple outer kinetochore components are phosphorylated by Aurora B kinase to activate the spindle assembly checkpoint (SAC) and to ensure accurate chromosome segregation. However, it is unknown whether Aurora B can phosphorylate inner kinetochore components to facilitate proper mitotic chromosome segregation. Here, we reported the structure of the fission yeast Schizosaccharomyces pombe Mis12–Nnf1 complex and showed that N-terminal residues 26–50 in Cnp3 (the CENP-C homolog of S. pombe) are responsible for interacting with the Mis12 complex. Interestingly, Thr28 of Cnp3 is a substrate of Ark1 (the Aurora B homolog of S. pombe), and phosphorylation impairs the interaction between the Cnp3 and Mis12 complex. The expression of a phosphorylation-mimicking Cnp3 mutant results in defective chromosome segregation due to improper kinetochore assembly. These results establish a previously uncharacterized regulatory mechanism involved in CENP-C–Mis12-facilitated kinetochore attachment error correction to ensure accurate chromosome segregation during mitosis.

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Aurora B Kinase Exists in a Complex with Survivin and INCENP and Its Kinase Activity Is Stimulated by Survivin Binding and Phosphorylation

Molecular Biology of the Cell ◽

10.1091/mbc.e02-02-0092 ◽

2002 ◽

Vol 13 (9) ◽

pp. 3064-3077 ◽

Cited By ~ 219

Author(s):

Margaret A. Bolton ◽

Weijie Lan ◽

Shannon E. Powers ◽

Mark L. McCleland ◽

Jian Kuang ◽

...

Keyword(s):

Cell Cycle ◽

Chromosome Segregation ◽

Kinase Activity ◽

Genetic Interactions ◽

Aurora B ◽

Hydrodynamic Properties ◽

Aurora B Kinase ◽

Survivin Gene ◽

Spindle Microtubules ◽

Cycle Dependent

Aurora B regulates chromosome segregation and cytokinesis and is the first protein to be implicated as a regulator of bipolar attachment of spindle microtubules to kinetochores. Evidence from several systems suggests that Aurora B is physically associated with inner centromere protein (INCENP) in mitosis and has genetic interactions with Survivin. It is unclear whether the Aurora B and INCENP interaction is cell cycle regulated and if Survivin physically interacts in this complex. In this study, we cloned theXenopus Survivin gene, examined its association with Aurora B and INCENP, and determined the effect of its binding on Aurora B kinase activity. We demonstrate that in the Xenopusearly embryo, all of the detectable Survivin is in a complex with both Aurora B and INCENP throughout the cell cycle. Survivin and Aurora B bind different domains on INCENP. Aurora B activity is stimulated >10-fold in mitotic extracts; this activation is phosphatase sensitive, and the binding of Survivin is required for full Aurora B activity. We also find the hydrodynamic properties of the Aurora B/Survivin/INCENP complex are cell cycle regulated. Our data indicate that Aurora B kinase activity is regulated by both Survivin binding and cell cycle-dependent phosphorylation.

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Modeling the temporal evolution of the spindle assembly checkpoint and role of Aurora B kinase

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.0810706106 ◽

2008 ◽

Vol 105 (51) ◽

pp. 20215-20220 ◽

Cited By ~ 37

Author(s):

H. B. Mistry ◽

D. E. MacCallum ◽

R. C. Jackson ◽

M. A. J. Chaplain ◽

F. A. Davidson

Keyword(s):

Spindle Assembly Checkpoint ◽

Temporal Evolution ◽

Spindle Assembly ◽

Aurora B ◽

Aurora B Kinase

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EB1 enables spindle microtubules to regulate centromeric recruitment of Aurora B

The Journal of Cell Biology ◽

10.1083/jcb.201307119 ◽

2014 ◽

Vol 204 (6) ◽

pp. 947-963 ◽

Cited By ~ 38

Author(s):

Budhaditya Banerjee ◽

Cortney A. Kestner ◽

P. Todd Stukenberg

Keyword(s):

Mitotic Chromosome ◽

Histone H3 ◽

Aurora B ◽

Dependent Manner ◽

Histone H2a ◽

Aurora B Kinase ◽

Chromosomal Passenger Complex ◽

Checkpoint Signaling ◽

Chromosomal Passenger ◽

Spindle Microtubules

The Aurora B kinase coordinates kinetochore–microtubule attachments with spindle checkpoint signaling on each mitotic chromosome. We find that EB1, a microtubule plus end–tracking protein, is required to enrich Aurora B at inner centromeres in a microtubule-dependent manner. This regulates phosphorylation of both kinetochore and chromatin substrates. EB1 regulates the histone phosphorylation marks (histone H2A phospho-Thr120 and histone H3 phospho-Thr3) that localize Aurora B. The chromosomal passenger complex containing Aurora B can be found on a subset of spindle microtubules that exist near prometaphase kinetochores, known as preformed K-fibers (kinetochore fibers). Our data suggest that EB1 enables the spindle microtubules to regulate the phosphorylation of kinetochores through recruitment of the Aurora B kinase.

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Aurora B kinase is recruited to multiple discrete kinetochore and centromere regions in human cells

The Journal of Cell Biology ◽

10.1083/jcb.201905144 ◽

2020 ◽

Vol 219 (3) ◽

Cited By ~ 16

Author(s):

Amanda J. Broad ◽

Keith F. DeLuca ◽

Jennifer G. DeLuca

Keyword(s):

Kinase Activity ◽

Critical Role ◽

Aurora B ◽

Histone H2a ◽

Aurora B Kinase ◽

Spindle Microtubules ◽

Kinetochore Region ◽

20 Nm ◽

Discrete Populations ◽

Early Mitosis

Aurora B kinase has a critical role in regulating attachments between kinetochores and spindle microtubules during mitosis. Early in mitosis, kinase activity at kinetochores is high to promote attachment turnover, and in later mitosis, activity decreases to ensure attachment stabilization. Aurora B localizes prominently to inner centromeres, and a population of the kinase is also detected at kinetochores. How Aurora B is recruited to and evicted from these regions to regulate kinetochore-microtubule attachments remains unclear. Here, we identified and investigated discrete populations of Aurora B at the centromere/kinetochore region. An inner centromere pool is recruited by Haspin phosphorylation of histone H3, and a kinetochore-proximal outer centromere pool is recruited by Bub1 phosphorylation of histone H2A. Finally, a third pool resides ~20 nm outside of the inner kinetochore protein CENP-C in early mitosis and does not require either the Bub1/pH2A/Sgo1 or Haspin/pH3 pathway for localization or activity. Our results suggest that distinct molecular pathways are responsible for Aurora B recruitment to centromeres and kinetochores.

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Mutations in orbit/mast reveal that the central spindle is comprised of two microtubule populations, those that initiate cleavage and those that propagate furrow ingression

The Journal of Cell Biology ◽

10.1083/jcb.200402052 ◽

2004 ◽

Vol 166 (1) ◽

pp. 49-60 ◽

Cited By ~ 92

Author(s):

Yoshihiro H. Inoue ◽

Matthew S. Savoian ◽

Takao Suzuki ◽

Endre Máthé ◽

Masa-Toshi Yamamoto ◽

...

Keyword(s):

Drosophila Melanogaster ◽

Time Lapse ◽

Aurora B ◽

Cleavage Furrow ◽

Imaging Studies ◽

Aurora B Kinase ◽

Central Spindle ◽

Spindle Microtubules ◽

Time Lapse Imaging ◽

Actin Rings

We address the relative roles of astral and central spindle microtubules (MTs) in cytokinesis of Drosophila melanogaster primary spermatocytes. Time-lapse imaging studies reveal that the central spindle is comprised of two MT populations, “interior” central spindle MTs found within the spindle envelope and “peripheral” astral MTs that probe the cytoplasm and initiate cleavage furrows where they contact the cortex and form overlapping bundles. The MT-associated protein Orbit/Mast/CLASP concentrates on interior rather than peripheral central spindle MTs. Interior MTs are preferentially affected in hypomorphic orbit mutants, and consequently the interior central spindle fails to form or is unstable. In contrast, peripheral MTs still probe the cortex and form regions of overlap that recruit the Pav-KLP motor and Aurora B kinase. orbit mutants have disorganized or incomplete anillin and actin rings, and although cleavage furrows initiate, they ultimately regress. Our work identifies a new function for Orbit/Mast/CLASP and identifies a novel MT population involved in cleavage furrow initiation.

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