Mitotic spindle association of TACC3 requires Aurora‐A‐dependent stabilization of a cryptic α‐helix

Burgess, Selena G.; Mukherjee, Manjeet; Sabir, Sarah; Joseph, Nimesh; Gutiérrez-Caballero, Cristina; Richards, Mark W.; Huguenin‐Dezot, Nicolas; Chin, Jason W.; Kennedy, Eileen J.; Pfuhl, Mark; Royle, Stephen; Gergely, Fanni; Bayliss, Richard

doi:10.15252/embj.201797902

Cited by 56 publications

(67 citation statements)

References 65 publications

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“…Our finding that homologues of human GTSE1 containing the important C-terminal domain exist in only vertebrates raises the question of whether the appearance of GTSE1 (and its interaction with CHC) may be evolutionarily linked to clathrin localization to the spindle and its MT-stabilizing function during mitosis. However, we find that TACC3 residues identified as required for interaction with CHC (Hood et al, 2013;Burgess et al, 2015Burgess et al, , 2018 are more distantly conserved, suggesting that a CHC-TACC3 complex on the spindle may have a more ancient origin (Fig. S3).…”

Section: Discussionmentioning

confidence: 92%

Clathrin’s adaptor interaction sites are repurposed to stabilize microtubules during mitosis

Rondelet

Lin

Singh

et al. 2020

Journal of Cell Biology

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Clathrin ensures mitotic spindle stability and efficient chromosome alignment, independently of its vesicle trafficking function. Although clathrin localizes to the mitotic spindle and kinetochore fiber microtubule bundles, the mechanisms by which clathrin stabilizes microtubules are unclear. We show that clathrin adaptor interaction sites on clathrin heavy chain (CHC) are repurposed during mitosis to directly recruit the microtubule-stabilizing protein GTSE1 to the spindle. Structural analyses reveal that these sites interact directly with clathrin-box motifs on GTSE1. Disruption of this interaction releases GTSE1 from spindles, causing defects in chromosome alignment. Surprisingly, this disruption destabilizes astral microtubules, but not kinetochore-microtubule attachments, and chromosome alignment defects are due to a failure of chromosome congression independent of kinetochore–microtubule attachment stability. GTSE1 recruited to the spindle by clathrin stabilizes microtubules by inhibiting the microtubule depolymerase MCAK. This work uncovers a novel role of clathrin adaptor-type interactions to stabilize nonkinetochore fiber microtubules to support chromosome congression, defining for the first time a repurposing of this endocytic interaction mechanism during mitosis.

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Section: Discussionmentioning

confidence: 92%

Clathrin’s adaptor interaction sites are repurposed to stabilize microtubules during mitosis

Rondelet

Lin

Singh

et al. 2020

Journal of Cell Biology

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“…Structural analysis of Aurora A confirms the importance of activation loop dynamics for activity and binding-partner interactions (Burgess et al, 2018). Phosphorylation, as well as TPX2 binding, stabilise the activation segement in an appropriate conformation for catalysis, and distinct Aurora A conformations can also be induced and/or stabilised by a huge variety of chemical small molecules (Dodson et al, 2010, McIntyre et al, 2017, Pitsawong et al, 2018, Tyler et al, 2007, Scutt et al, 2009, Damodaran et al, 2017.…”

Section: Introductionmentioning

confidence: 83%

“…Aurora C is most highly expressed in germ cells, where it can functionally replace Auora B as a chromosome passenger protein (Carmena and Earnshaw, 2003). Aurora A biology is controlled spatially via targeted subcellular localization mediated through the formation of dynamic protein complexes with non-catalytic binding partners such as TPX2 , Bayliss et al, 2003, Wittmann et al, 2000, TACC3 (Burgess et al, 2018) and NMYC (Richards et al, 2016), which control distinct Aurora A signaling outputs relevant to cellular dynamics. Indeed, Aurora A/TPX2 holoenzyme complexes co-localise at the polar end of spindle microtubules, where Aurora A is maintained in dynamic pools alongside the mitoitc kinesin Eg5, which is localised by a C-terminal region of TPX2 (Eckerdt et al, 2008, Zeng et al, 2010.…”

Section: Introductionmentioning

confidence: 99%

Covalent Aurora A regulation by the metabolic integrator coenzyme A

Tsuchiya

Byrne

Burgess

et al. 2018

Preprint

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Aurora A is a cell cycle protein kinase implicated in multiple human cancers, and several Aurora Aspecific kinase inhibitors have progressed into clinical trials. In this study, we report structural and cellular analysis of a novel biochemical mode of Aurora A inhibition, which occurs through reversible covalent interaction with the universal metabolic integrator coenzyme A (CoA). Mechanistically, the CoA 3'-phospho ADP moiety interacts with Thr 217, an Aurora A selectivity filter, which permits the formation of an unprecedented covalent bond with Cys 290 in the kinase activation segment, lying some 15 Å away. CoA modification (CoAlation) of endogenous Aurora A is rapidly induced by oxidative stresses at Cys 290 in human cells, and microinjection of CoA into mouse embryos perturbs meitoic spindle formation and chromosome alignment. Aurora A regulation by CoA reveals how targeting of Aurora A might be accomplished in the future by development of a 'doubleanchored' covalent inhibitor.

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“…Formation of this complex (hereafter referred to as the CHC/TACC3 complex) and its recruitment to spindles depends on the direct interaction between clathrin heavy chain (CHC) and TACC3 phosphorylated on serine S558 by the Aurora A kinase, thereby restricting the function of this clathrin complex on microtubules to mitosis. (Hubner et al, 2010;Fu et al, 2010;Lin et al, 2010;Booth et al, 2011;Burgess et al, 2018;Hood et al, 2013;Burgess et al, 2015). While the recruitment of the CHC/TACC3 complex during mitosis to spindles is necessary for MT stabilization and chromosome alignment, the mechanisms by which it stabilizes MTs remains unclear.…”

Section: Introductionmentioning

confidence: 99%

Clathrin’s adaptor interaction sites are repurposed to stabilize microtubules during mitosis

Rondelet

Lin

Singh

et al. 2019

Preprint

View full text Add to dashboard Cite

Clathrin plays an important role to ensure mitotic spindle stability and efficient chromosome alignment, independently of its well-characterized functions in vesicle trafficking. While clathrin clearly localizes to the mitotic spindle and kinetochore-fiber microtubule bundles, the mechanisms by which clathrin stabilizes microtubules have remained elusive. Here we show that clathrin adaptor interaction sites on clathrin heavy chain (CHC) are repurposed during mitosis to directly recruit the microtubule-stabilizing protein GTSE1 to the mitotic spindle.Structural analyses reveal that multiple clathrin-box motifs on GTSE1 interact directly with different clathrin adaptor interaction sites on CHC, in a manner structurally analogous to that which occurs between adaptor proteins and CHC near membranes. Specific disruption of this interaction in cells releases GTSE1 from spindles and causes defects in chromosome alignment. Surprisingly, this disruption causes destabilization of astral microtubules, but not kinetochore-microtubule attachments, and the resulting chromosome alignment defect is due to a failure of chromosome congression independent of kinetochore-microtubule attachment stability. Finally, we show that GTSE1 recruited to the spindle by clathrin stabilizes 2 microtubules and promotes chromosome congression by inhibiting the activity of the microtubule depolymerase MCAK. This work thus uncovers a novel role of clathrin to stabilize non-kinetochore-fiber microtubules to support chromosome congression. This role is carried out via clathrin adaptor-type interactions of CHC with GTSE1, defining for the first time an important repurposing of this endocytic interaction mechanism during mitosis. Rizk et al., 2009; Walczak et al., 1996;Wordeman and Mitchison, 1995) (Moore et al., 2005). It was proposed that a function of TACC3 is to antagonize MCAK's potent activity at centrosomes to stabilize astral MTs, presumably by promoting activity of the microtubule depolymerase XMAP215/ch-Tog (Kinoshita et al., 2005). However, whether clathrin (or the CHC/TACC3 complex) is required for astral microtubule stabilization has not been addressed.Another potential functional member of the CHC/TACC3 complex is GTSE1 (pronounced "jitsee-one" (Monte et al., 2000)), an intrinsically disordered protein that has been shown by proteomic analysis to interact with the CHC/TACC3 complex in cells (Hubner et al., 2010). Two recent studies showed that GTSE1 localizes to the spindle during mitosis, and like TACC3, is necessary for astral microtubule stabilization and efficient chromosome alignment (Bendre et al., 2016;Tipton et al., 2017). We showed that GTSE1 stabilizes MTs in mitosis by inhibiting MCAK activity (Bendre et al., 2016). In that study, cancer cells either depleted or knocked-out of GTSE1 showed destabilization of both astral MTs and kinetochore-MT attachments.Another study reported that a specific "slow turnover" population of MTs become stabilized upon depletion of GTSE1(Tipton et al., 2017). The reason for these seemingly contr...

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Mitotic spindle association of TACC3 requires Aurora‐A‐dependent stabilization of a cryptic α‐helix

Cited by 56 publications

References 65 publications

Clathrin’s adaptor interaction sites are repurposed to stabilize microtubules during mitosis

Clathrin’s adaptor interaction sites are repurposed to stabilize microtubules during mitosis

Covalent Aurora A regulation by the metabolic integrator coenzyme A

Clathrin’s adaptor interaction sites are repurposed to stabilize microtubules during mitosis

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