A small-molecule inhibitor of Haspin alters the kinetochore functions of Aurora B

A, De Antoni; Maffini, Stefano; Knapp, Stefan; Musacchio, Andrea; Santaguida, Stefano

doi:10.1083/jcb.201205119

Cited by 102 publications

(140 citation statements)

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“…4B, supplemental Table S7). The inhibition of RSK is unlikely to be caused by off-target 5-ITu inhibition, because the concentration of the inhibitor used in this study does not significantly affect RSK kinases activity in vitro (1 M 5-ITu reduces only of the 3% RSKs kinase activities) (11). Therefore, we consider the down-regulation of predicted RSK substrates as an indirect effect of Haspin inhibition.…”

Section: Kinase-substrate Enrichment Analysis (Ksea) Reveals Inactivamentioning

confidence: 99%

“…1C) was not affected. This was probably because of incomplete Aurora B delocalization from the centromeres (9,11,12,39).…”

Section: Generation Of a Data Set Of Haspin Modulated Chromatin Assomentioning

confidence: 99%

“…CLKs are dual specificity protein kinases involved in pre-mRNA processing (52). Inhibition of CLK family kinases can be interpreted as a direct effect of 5-ITu treatment as reported by a specificity screen showing that 5-ITu inhibits in vitro Clk2 (94% of the kinase activity) and less efficiently Clk3 (50% of the kinase activity) kinases (11). Eighteen phosphorylation sites were predicted as CLK substrates; more than 50% of those were from proteins involved in gene expression and splicing mechanisms (Fig.…”

Section: Kinase-substrate Enrichment Analysis (Ksea) Reveals Inactivamentioning

confidence: 99%

“…S1) to inhibit Haspin kinase activity and to study the effect of its inhibition in human mitotic HeLa cells. A specificity screen performed on a panel of 138 kinases in vitro revealed the selectivity of 5-ITu toward Haspin and the members of Clk kinase family (11). To confirm efficient Haspin inhibition in cells, we monitored the phosphorylation of Thr 3 of histone H3 by IF (supplemental Fig.…”

Section: Generation Of a Data Set Of Haspin Modulated Chromatin Assomentioning

confidence: 99%

“…H3Thr 3 phosphorylation (H3Thr 3ph ) is required for the localization of Aurora B at the centromere (8 -10). Inactivation of Haspin catalytic activity by ATP mimetic inhibitors induces Aurora B centromeric delocalization, leading to a loss of phosphorylation in chromatin associated Aurora B substrates (11,12). To date, apart from this well-characterized centromeric function of Haspin activity, the broader cellular functions of the kinase and the phosphorylation events that control these remain essentially unknown.…”

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confidence: 99%

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Modulation of the Chromatin Phosphoproteome by the Haspin Protein Kinase

Maiolica

Medina-Redondo

Schoof

et al. 2014

Molecular & Cellular Proteomics

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Recent discoveries have highlighted the importance of Haspin kinase activity for the correct positioning of the kinase Aurora B at the centromere. Haspin phosphorylates Thr 3 of the histone H3 (H3), which provides a signal for Aurora B to localize to the centromere of mitotic chromosomes. To date, histone H3 is the only confirmed Haspin substrate. We used a combination of biochemical, pharmacological, and mass spectrometric approaches to study the consequences of Haspin inhibition in mitotic cells. We quantified 3964 phosphorylation sites on chromatin-associated proteins and identified a Haspin protein-protein interaction network. We determined the Haspin consensus motif and the co-crystal structure of the kinase with the histone H3 tail.

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Section: Kinase-substrate Enrichment Analysis (Ksea) Reveals Inactivamentioning

confidence: 99%

“…1C) was not affected. This was probably because of incomplete Aurora B delocalization from the centromeres (9,11,12,39).…”

Section: Generation Of a Data Set Of Haspin Modulated Chromatin Assomentioning

confidence: 99%

Section: Kinase-substrate Enrichment Analysis (Ksea) Reveals Inactivamentioning

confidence: 99%

Section: Generation Of a Data Set Of Haspin Modulated Chromatin Assomentioning

confidence: 99%

mentioning

confidence: 99%

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Modulation of the Chromatin Phosphoproteome by the Haspin Protein Kinase

Maiolica

Medina-Redondo

Schoof

et al. 2014

Molecular & Cellular Proteomics

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N₁‐Benzoylated 5‐(4‐pyridinyl)indazole‐based kinase inhibitors: Attaining haspin and Clk4 selectivity via modulation of the benzoyl substituents

Aboelfotouh,

Abdallah,

Khalifa

et al. 2024

Archiv der Pharmazie

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Haspin and Clk4 are both understudied protein kinases (PKs), offering potential targets for the development of new anticancer agents. Thus, the identification of new inhibitors targeting these PKs is of high interest. However, the inhibitors targeting haspin or Clk4 developed to date show a poor selectivity profile over other closely related PKs, increasing the risk of side effects. Herein, we present two newly developed N1‐benzyolated 5‐(4‐pyridinyl)indazole‐based inhibitors (18 and 19), derived from a newly identified indazole hit. These inhibitors exhibit an exceptional inhibitory profile toward haspin and/or Clk4. Compound 18 (2‐acetyl benzoyl) showed a preference to inhibit Clk4 and haspin over a panel of closely related kinases, with sixfold selectivity for Clk4 (IC50 = 0.088 and 0.542 μM, respectively). Compound 19 (4‐acetyl benzoyl) showed high selectivity against haspin over the common off‐target kinases (Dyrks and Clks) with an IC50 of 0.155 μM for haspin. Molecular docking studies explained the remarkable selectivity of 18 and 19, elucidating how the new scaffold can be modified to toggle between inhibition of haspin or Clk4, despite the high homology of the ATP‐binding sites. Their distinguished profile allows these compounds to be marked as interesting chemical probes to assess the selective inhibition of haspin and/or Clk4.

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CRISPR/Cas9 screening identifies a kinetochore‐microtubule dependent mechanism for Aurora‐A inhibitor resistance in breast cancer

Chen

Wen

Liu

et al. 2021

Cancer Communications

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Background: Overexpression of Aurora-A (AURKA) is a feature of breast cancer and associates with adverse prognosis. The selective Aurora-A inhibitor alisertib (MLN8237) has recently demonstrated promising antitumor responses as a single agent in various cancer types but its phase III clinical trial was reported as a failure since MLN8237 did not show an apparent effect in prolonging the survival of patients. Thus, identification of potential targets that could enhance the activity of MLN8237 would provide a rationale for drug combination to achieve better therapeutic outcome. Methods: Here, we conducted a systematic synthetic lethality CRISPR/Cas9 screening of 507 kinases using MLN8237 in breast cancer cells and identified a number of targetable kinases that displayed synthetic lethality interactions with MLN8237. Then, we performed competitive growth assays, colony formation assays, cell viability assays, apoptosis assays, and xenograft murine model to Abbreviations: AURKA/Aurora-A, aurora kinase A; AURKB/Aurora-B, aurora kinase B; BUB1B, BUB1 mitotic checkpoint serine/threonine kinase B; CHEK1, checkpoint kinase 1; CPC, chromosomal passenger complex; CRISPR, clustered regularly interspaced short palindromic repeats; CSNK1A1, casein kinase 1 alpha 1; GSG2, germ cell-specific gene 2; DAPI, 4′,6-diamidino-2-phenylindole; DDR1, discoidin domain receptor tyrosine kinase 1; EGFP, enhanced green fluorescent protein; EGFP-sgCtrl, cells labeled with EGFP and introduced with a non-targeting sgRNA; ExMD, extends mitotic duration; K-fibers, kinetochore fibers; KMN network, kinetochore null protein 1 (KNL1)-missegregation 12 (MIS12) complex-nuclear division cycle 80 (NDC80) complex; KT, kinetochore; KT-MT, kinetochore-microtubule attachment; MAGeCK, model-based analysis of genome-wide CRISPR/Cas9 knockout; MCAK, mitotic centromere-associated kinesin; mCherry-sgCtrl, cells labeled with mCherry and introduced with a non-targeting sgRNA; mCherry-sgGSG2, cells labeled with mCherry and introduced with a sgRNA targeting GSG2; MINK1, misshapen like kinase 1; MOI, multiplicity of infection; MT, microtubule; NEK1, NIMA related kinase 1; NGS, next-generation sequencing; PCA, principal component analysis; PCM, pericentriolar material; pH3T3, phosphorylated threonine 3 of histone H3; QC, quality control; RB1, retinoblastoma; sgRNAs, single guide RNAs; TCGA, The Cancer Genome Atlas This is an open access article under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

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A small-molecule inhibitor of Haspin alters the kinetochore functions of Aurora B

Abstract: A chemical biology study characterizes the role of Haspin kinase in centromere recruitment of the chromosome passenger complex and in spindle assembly checkpoint function.

Cited by 102 publications

References 95 publications

Modulation of the Chromatin Phosphoproteome by the Haspin Protein Kinase

Modulation of the Chromatin Phosphoproteome by the Haspin Protein Kinase

N₁‐Benzoylated 5‐(4‐pyridinyl)indazole‐based kinase inhibitors: Attaining haspin and Clk4 selectivity via modulation of the benzoyl substituents

CRISPR/Cas9 screening identifies a kinetochore‐microtubule dependent mechanism for Aurora‐A inhibitor resistance in breast cancer

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A small-molecule inhibitor of Haspin alters the kinetochore functions of Aurora B

Abstract: A chemical biology study characterizes the role of Haspin kinase in centromere recruitment of the chromosome passenger complex and in spindle assembly checkpoint function.

Cited by 102 publications

References 95 publications

Modulation of the Chromatin Phosphoproteome by the Haspin Protein Kinase

Modulation of the Chromatin Phosphoproteome by the Haspin Protein Kinase

N1‐Benzoylated 5‐(4‐pyridinyl)indazole‐based kinase inhibitors: Attaining haspin and Clk4 selectivity via modulation of the benzoyl substituents

CRISPR/Cas9 screening identifies a kinetochore‐microtubule dependent mechanism for Aurora‐A inhibitor resistance in breast cancer

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About

N₁‐Benzoylated 5‐(4‐pyridinyl)indazole‐based kinase inhibitors: Attaining haspin and Clk4 selectivity via modulation of the benzoyl substituents