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

Maeda, Katsutoshi; Yoneda, Mitsuhiro; Nakagawa, Takeya; Ikeda, Kazuhiro; Higashi, Miki; Nakagawa, Kaori; Miyakoda, Mana; Yui, Katsuyuki; Oda, Hiroaki; Inoue, Satoshi; Ito, Takashi

doi:10.1111/gtc.12630

Cited by 10 publications

(3 citation statements)

References 18 publications

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“…In yeast, the corresponding S121 is phosphorylated by Bub1 to recruit shugoshin, an important step for centromere function in chromosome segregation [58][59][60]. This serine is replaced by threonine (T120) in human H2A and H2A.X, and its phosphorylation is associated with transcriptional activation and mitotic chromosome segregation [61][62][63]. Consistent with these data, we detected only a few events of S129 phosphorylation in H2A.W in samples originating from leaves or seedlings that contain low amounts of dividing cells.…”

Section: Discussionsupporting

confidence: 85%

Crosstalk between H2A variant-specific modifications impacts vital cell functions

Schmuecker

Lei

Lorković

et al. 2021

Preprint

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Histone variants are distinguished by specific substitutions and motifs that might be subject to post-translational modifications (PTMs). Compared with the high conservation of H3 variants, the N- and C-terminal tails of H2A variants are more divergent and are potential substrates for a more complex array of PTMs, which have remained largely unexplored. We used mass spectrometry to inventory the PTMs of the two heterochromatin-enriched variants H2A.W.6 and H2A.W.7 of Arabidopsis, which harbor the C-terminal motif KSPK. This motif is also found in macroH2A variants in animals and confers specific properties to the nucleosome. We showed that H2A.W.6 is phosphorylated by the cell cycle-dependent kinase CDKA specifically at KSPK. In contrast, this modification is absent on H2A.W.7, which also harbors the SQ motif associated with the variant H2A.X. Phosphorylation of the SQ motif is critical for the DNA damage response but is suppressed in H2A.W.7 by phosphorylation of KSPK. To identify factors involved in this suppression mechanism, we performed a synthetic screen in fission yeast expressing a mimic of the Arabidopsis H2A.W.7. Among those factors was the BRCT-domain protein Mdb1. We showed that phosphorylation of KSPK prevents binding of the BRCT-domain protein Mdb1 to phosphorylated SQ and as a result hampers response to DNA damage. Hence, cross-talks between motif-specific PTMs interfere with the vital functions of H2A variants. Such interference could be responsible for the mutual exclusion of specific motifs between distinct H2A variants. We conclude that sequence innovations in H2A variants have potentiated the acquisition of many specific PTMs with still unknown functions. These add a layer of complexity to the nucleosome properties and their impact in chromatin regulation.

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Section: Discussionsupporting

confidence: 85%

Crosstalk between H2A variant-specific modifications impacts vital cell functions

Schmuecker

Lei

Lorković

et al. 2021

Preprint

View full text Add to dashboard Cite

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“…Moreover, neither mutation nor depletion of NHK-1 prevents mitotic H2AT119ph in flies or Drosophila S2 cells 86 , 88 . Instead, the kinetochore-binding kinase Bub1 is clearly vital for H2AT120ph in budding and fission yeast, mice, and human cells 18 , 89 , 90 . Together, there is little evidence to support a significant role for VRK1 in direct phosphorylation of H3T3ph or other histone residues in mitosis.…”

Section: Discussionmentioning

confidence: 99%

Dissecting the roles of Haspin and VRK1 in histone H3 phosphorylation during mitosis

Cartwright

Harris

Meyer

et al. 2022

Sci Rep

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Protein kinases that phosphorylate histones are ideally-placed to influence the behavior of chromosomes during cell division. Indeed, a number of conserved histone phosphorylation events occur prominently during mitosis and meiosis in most eukaryotes, including on histone H3 at threonine-3 (H3T3ph). At least two kinases, Haspin and VRK1 (NHK-1/ballchen in Drosophila), have been proposed to carry out this modification. Phosphorylation of H3 by Haspin has defined roles in mitosis, but the significance of VRK1 activity towards histones in dividing cells has been unclear. Here, using in vitro kinase assays, KiPIK screening, RNA interference, and CRISPR/Cas9 approaches, we were unable to substantiate a direct role for VRK1, or its paralogue VRK2, in the phosphorylation of threonine-3 or serine-10 of Histone H3 in mitosis, although loss of VRK1 did slow cell proliferation. We conclude that the role of VRKs, and their more recently identified association with neuromuscular disease and importance in cancers of the nervous system, are unlikely to involve mitotic histone kinase activity. In contrast, Haspin is required to generate H3T3ph during mitosis.

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“…Moreover, neither mutation nor depletion of NHK-1 prevents mitotic H2AT119ph in flies or Drosophila S2 cells (70,74). Instead, the kinetochore-binding kinase Bub1 is clearly vital for H2AT120ph in budding and fission yeast, mice, and human cells (75-77). Together, there is little evidence to support a significant role for VRK1 in direct phosphorylation of H3T3ph or other histone residues in mitosis.…”

Section: Discussionmentioning

confidence: 99%

Dissecting the roles of Haspin and VRK1 in Histone H3 threonine-3 phosphorylation during mitosis

Cartwright

Harris

Meyer

et al. 2021

Preprint

View full text Add to dashboard Cite

Protein kinases that phosphorylate histones are ideally-placed to influence the behavior of chromosomes during cell division. Indeed, a number of conserved histone phosphorylation events occur prominently during mitosis and meiosis in most eukaryotes, including on histone H3 at threonine-3 (H3T3ph). At least two kinases, Haspin and VRK1 (NHK-1/ballchen in Drosophila), have been proposed to carry out this modification. Phosphorylation of H3 by Haspin has defined roles in mitosis, but the significance of VRK1 activity towards histones in dividing cells has been unclear. Here, using in vitro kinase assays, KiPIK screening, RNA interference, and CRISPR/Cas9 approaches, we were unable to substantiate a direct role for VRK1, or its homologue VRK2, in the phosphorylation of threonine-3 or serine-10 of Histone H3 in mitosis, although loss of VRK1 did slow cell proliferation. We conclude that the role of VRK1, and its more recently identified association with neuromuscular disease in humans, is unlikely to involve mitotic histone kinase activity. In contrast, Haspin is required to generate H3T3ph during mitosis.

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

Cited by 10 publications

References 18 publications

Crosstalk between H2A variant-specific modifications impacts vital cell functions

Crosstalk between H2A variant-specific modifications impacts vital cell functions

Dissecting the roles of Haspin and VRK1 in histone H3 phosphorylation during mitosis

Dissecting the roles of Haspin and VRK1 in Histone H3 threonine-3 phosphorylation during mitosis

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