Probing the catalytic functions of Bub1 kinase using the small molecule inhibitors BAY-320 and BAY-524

Baron, Anna P.; Schubert, Conrad von; Cubizolles, Fabien; Siemeister, Gerhard; Hitchcock, Marion; Mengel, Anne; Schröder, Jörg; Fernández-Montalván, Amaury E.; Nussbaum, Franz von; Mumberg, Dominik; Nigg, Erich A.

doi:10.7554/elife.12187

Cited by 60 publications

(112 citation statements)

References 106 publications

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“…Extremely shortened mitosis induces massive chromosome missegregation and/or tetraploidization that elevates cell stress beyond the lethal level [164,165]. In line with the idea, inhibitors of Mps1 kinase were shown to exert anti-tumor activities [166][167][168], and anti-SAC compounds were reported to sensitize cancer cells to microtubule-targeting drugs [169][170][171][172].…”

Section: Accepted Manuscriptmentioning

confidence: 94%

Chromosomal instability: A common feature and a therapeutic target of cancer

Tanaka

Hirota

2016

Biochimica et Biophysica Acta (BBA) - Reviews on Cancer

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Section: Accepted Manuscriptmentioning

confidence: 94%

Chromosomal instability: A common feature and a therapeutic target of cancer

Tanaka

Hirota

2016

Biochimica et Biophysica Acta (BBA) - Reviews on Cancer

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“…We performed iterated sensitive homology searches with jackhammer (Finn et al, 2011) (based on the TPR, kinase, CDI, GLEBS and KEN boxes) using a permissive E-value and bitscore cut-off to include diverged homologs on uniprot release 2016_08 and Ensemble Genomes 32 (http://www.ebi.ac.uk/Tools/hmmer/search/jackhmmer). Incompletely predicted genes were searched against whole genome shotgun contigs (wgs, http://www.ncbi.nlm.nih.gov/genbank/wgs) using tblastn.…”

Section: Methodsmentioning

confidence: 99%

Phylogenomics-guided discovery of a novel conserved cassette of short linear motifs in BubR1 essential for the spindle checkpoint

Tromer

Bade

Snel

et al. 2016

Preprint

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The spindle assembly checkpoint (SAC) maintains genomic integrity by preventing progression of mitotic cell division until all chromosomes are stably attached to spindle microtubules. The SAC critically relies on the paralogues Bub1 and BubR1/Mad3, which integrate kinetochore-spindle attachment status with generation of the anaphase inhibitory complex MCC. We previously reported on the widespread occurrences of independent gene duplications of an ancestral 'MadBub' gene in eukaryotic evolution and the striking parallel subfunctionalization that lead to loss of kinase function in BubR1/Mad3-like paralogues. Here, we present an elaborate subfunctionalization analysis of the Bub1/BubR1 gene family and perform de novo sequence discovery in a comparative phylogenomics framework to trace the distribution of ancestral sequence features to extant paralogues throughout the eukaryotic tree of life. We show that known ancestral sequence features are consistently retained in the same functional paralogue: GLEBS/CMI/ CDII/kinase in the Bub1-like and KEN1/KEN2/D-Box in the BubR1/ Mad3-like. The recently described ABBA motif can be found in either or both paralogues. We however discovered two additional ABBA motifs that flank KEN2. This cassette of ABBA1-KEN2-ABBA2 forms a strictly conserved module in all ancestral and BubR1/Mad3-like proteins, suggestive of a specific and crucial SAC function. Indeed, deletion of the ABBA motifs in human BUBR1 abrogates the SAC and affects APC/C-Cdc20 interactions. Our detailed comparative genomics analyses thus enabled discovery of a conserved cassette of motifs essential for the SAC and shows how this approach can be used to uncover hitherto unrecognized functional protein features.

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“…It was shown that Bub1 phosphorylates the conserved histone H2A serine 121, which corresponds to H2A threonine 120 in humans. The histone H2A S121A mutant, in which all cellular H2A serine 121 is replaced by alanine, mimics the Bub1 kinase-dead mutant (Bub1-KD) in perturbing the centromeric localization of the Shugoshin protein, which is also associated with the pericentromeric chromosomal region and plays multiple roles in ensuring the accuracy of chromosome segregation during both mitosis and meiosis (Asghar et al, 2015;Baron et al, 2016;Lin, Jia, Tomchick, Luo, & Yu, 2014;Liu, Jia, & Yu, 2013;Liu et al, 2015;Marston, 2015). In addition to its role in histone H2A phosphorylation, it has also been reported that over-expression of Bub1 drives chromosomal instability through hyperactivation of Aurora B kinase (Ricke, Jeganathan, & van Deursen, 2011).…”

Section: Introductionmentioning

confidence: 99%

Defects in centromeric/pericentromeric histone H2A T120 phosphorylation by hBUB1 cause chromosome missegregation producing multinucleated cells

et al. 2018

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Histone H2A phosphorylation plays a role both in chromatin condensation during mitosis and in transcriptional activation during the G1/S transition. Bub1 and NHK1/VRK1 have been identified as histone H2A kinases. However, little is known about the importance of histone H2A phosphorylation in chromosome segregation. Here, we expressed recombinant hBUB1 and confirmed that it phosphorylates histone H2A T120 in the in vitro-assembled nucleosome. Knockdown (KD) of BUB1 decreases bulk H2A T120 phosphorylation in HeLa cells, whereas hBUB1 is upregulated during mitosis, which corresponds with H2A T120 phosphorylation. ChIP-qPCR of the DXZ1 centromeric and γ-ALR pericentromeric region showed that BUB1 localizes to this region and increases local H2A T120 phosphorylation during M phase. BUB1 KD did not induce apoptosis but increased the M phase cell population, as detected by flow cytometry. BUB1 KD also caused an abnormal metaphase and telophase, resulting in multinucleated cells and impaired cancer cell growth both in vitro and in vivo. Over-expression of the histone H2A T120D or T120E mutations, which mimic phosphorylated threonine, decreased the number of multinucleated cells caused by BUB1 KD. These results strengthen the apparent importance of BUB1-mediated H2A T120 phosphorylation in normal mitosis.

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Probing the catalytic functions of Bub1 kinase using the small molecule inhibitors BAY-320 and BAY-524

Cited by 60 publications

References 106 publications

Chromosomal instability: A common feature and a therapeutic target of cancer

Chromosomal instability: A common feature and a therapeutic target of cancer

Phylogenomics-guided discovery of a novel conserved cassette of short linear motifs in BubR1 essential for the spindle checkpoint

Defects in centromeric/pericentromeric histone H2A T120 phosphorylation by hBUB1 cause chromosome missegregation producing multinucleated cells

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