Dai, J., Sultan, S., Taylor, S.S. & Higgins, J.M. The kinase haspin is required for mitotic histone H3 Thr 3 phosphorylation and normal metaphase chromosome alignment. Genes Dev. 19, 472-488

Division of Rheumatology, Immunology and Allergy, Department of Medicine, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
Genes & Development (Impact Factor: 10.8). 03/2005; 19(4):472-88. DOI: 10.1101/gad.1267105
Source: PubMed


Post-translational modifications of conserved N-terminal tail residues in histones regulate many aspects of chromosome activity. Thr 3 of histone H3 is highly conserved, but the significance of its phosphorylation is unclear, and the identity of the corresponding kinase unknown. Immunostaining with phospho-specific antibodies in mammalian cells reveals mitotic phosphorylation of H3 Thr 3 in prophase and its dephosphorylation during anaphase. Furthermore we find that haspin, a member of a distinctive group of protein kinases present in diverse eukaryotes, phosphorylates H3 at Thr 3 in vitro. Importantly, depletion of haspin by RNA interference reveals that this kinase is required for H3 Thr 3 phosphorylation in mitotic cells. In addition to its chromosomal association, haspin is found at the centrosomes and spindle during mitosis. Haspin RNA interference causes misalignment of metaphase chromosomes, and overexpression delays progression through early mitosis. This work reveals a new kinase involved in composing the histone code and adds haspin to the select group of kinases that integrate regulation of chromosome and spindle function during mitosis and meiosis.

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Available from: Stephen S Taylor, Oct 07, 2015
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    • "Thus, in addition to H3K4me3, the phosphorylation of T3 and T6, but not S10, probably acts as additional ''on/off switches'' (Fischle et al., 2003) to control the sites at which Dnmt3a associates with chromatin. H3T3ph appears exclusively during mitosis (Dai et al., 2005; Markaki et al., 2009) and H3T6ph occurs mainly during interphase (Garske et al., 2010; Metzger et al., 2010). "
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    ABSTRACT: Histone modification and DNA methylation are associated with varying epigenetic "landscapes," but detailed mechanistic and functional links between the two remain unclear. Using the ATRX-DNMT3-DNMT3L (ADD) domain of the DNA methyltransferase Dnmt3a as a paradigm, we apply protein engineering to dissect the molecular interactions underlying the recruitment of this enzyme to specific regions of chromatin in mouse embryonic stem cells (ESCs). By rendering the ADD domain insensitive to histone modification, specifically H3K4 methylation or H3T3 phosphorylation, we demonstrate the consequence of dysregulated Dnmt3a binding and activity. Targeting of a Dnmt3a mutant to H3K4me3 promoters decreases gene expression in a subset of developmental genes and alters ESC differentiation, whereas aberrant binding of another mutant to H3T3ph during mitosis promotes chromosome instability. Our studies support the general view that histone modification "reading" and DNA methylation are closely coupled in mammalian cells, and suggest an avenue for the functional assessment of chromatin-associated proteins. Copyright © 2015 Elsevier Inc. All rights reserved.
    Molecular cell 06/2015; 59(1). DOI:10.1016/j.molcel.2015.05.017 · 14.02 Impact Factor
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    • "In agreement with its function in cell division, Haspin localizes at the chromosomes during mitosis (Dai et al. 2005; Ashtiyani et al. 2011; Kurihara et al. 2011). Dai et al. (2005) demonstrated that reduced levels of H3T3 phosphorylation induced by RNA interference-mediated depletion of the Haspin kinase, results in late-prometaphase configurations in mammalian cells. Overexpression of Haspin, on the other hand, causes a deficit in cell proliferation and a delay in the transition from G2 to mitosis. "
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    ABSTRACT: Post-translational histone modifications, such as methylation and phosphorylation, play an important role in determining chromatin states associated with gene activation or repression. Histone H3 phosphorylation in particular has been linked to a variety of cellular processes during the cell cycle. H3 phosphorylation is involved in chromosome condensation and segregation during mitosis and meiosis in plants and animals. During interphase, H3 phosphorylation has been implicated in transcriptional regulation, DNA replication and apoptosis. Phosphorylation also occurs in the histone variants, H3.3 and CENH3, during cell division. The diverse and sometimes contrasting processes in which H3 phosphorylation participates have made difficult to completely understand its function. In addition, functional differences on H3 phosphorylation have been observed in diverse organisms despite the conservation of the modified residue. Here we discuss the most recent findings about the roles of histone H3 phosphorylation, the proteins involved in phosphorylating particular residues and the mechanisms by which this modification results in a particular gene expression state. The differences and similarities between plants and other model systems are emphasized.
    Epigenetics in Plants of Agronomic Importance: Fundamentals and Applications, 09/2014: chapter 4: pages 47-70; Springer International Publishing.
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    • "Although the dephosphorylation of H3T3ph at the exit from M phase is required for proper chromosome decondensation and nuclear envelope formation (Kelly et al., 2010), the molecular mechanisms that limit H3T3ph to M phase remain unclear. Mitotic H3T3 phosphorylation is catalyzed by Haspin (Dai et al., 2005), which is an atypical protein kinase in several regards . For example, in most kinases, the highly conserved DFG (Asp-Phe-Gly) motif anchors the N-terminal portion of the activation segment and coordinates catalytic magnesium (Nolen et al., 2004), but, in Haspin, it is changed into DYT (Asp-Tyr-Thr). "
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    ABSTRACT: The mitosis-specific phosphorylation of histone H3 at Thr3 (H3T3ph) plays an important role in chromosome segregation by recruiting Aurora B. H3T3 phosphorylation is catalyzed by Haspin, an atypical protein kinase whose kinase domain is intrinsically active without phosphorylation at the activation loop. Here, we report the molecular basis for Haspin inhibition during interphase and its reactivation in M phase. We identify a conserved basic segment that autoinhibits Haspin during interphase. This autoinhibition is neutralized when Cdk1 phosphorylates the N terminus of Haspin in order to recruit Polo-like kinase (Plk1/Plx1), which, in turn, further phosphorylates multiple sites at the Haspin N terminus. Although Plx1, and not Aurora B, is critical for H3T3 phosphorylation in Xenopus egg extracts, Plk1 and Aurora B both promote this modification in human cells. Thus, M phase-specific H3T3 phosphorylation is governed by the combinatorial action of mitotic kinases that neutralizes Haspin autoinhibition through a mechanism dependent on multisite phosphorylation.
    Molecular cell 10/2013; 52(5). DOI:10.1016/j.molcel.2013.10.002 · 14.02 Impact Factor
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