Genomes and Genes
Function of the chromosomal kinase haspin in mitosis
Principal Investigator: JONATHAN HIGGINS
Abstract: We have recently discovered a novel mitotic histone kinase, haspin, that has homologs in diverse eukaryotes. Haspin phosphorylates Thr-3 in the N-terminal tail of histone H3. In human cells, H3 Thr-3 phosphorylation is first detected on chromosome arms in late G2/early prophase, becomes focused at centromeres by prometaphase, and declines during anaphase. In vitro, haspin specifically phosphorylates histone H3 at Thr-3, and depletion of haspin by RNA interference (RNAi) reveals that it is required for H3 Thr-3 phosphorylation in mitotic cells. Haspin associates with condensed chromosomes, particularly at centromeres, and is also found at the centrosomes during mitosis. Importantly, haspin RNAi causes misalignment of metaphase chromosomes and spindle defects, and overexpression delays progression through early mitosis. Our more recent data suggest that haspin is required for the maintenance of sister chromatid cohesion and centromeric aurora B localization prior to anaphase. We have also isolated candidate haspin-binding proteins that are consistent with haspin function at the centrosome and spindle. This work reveals a new enzyme involved in composing the histone code and adds haspin to the select group of kinases that regulate chromosome dynamics and spindle activity during mitosis. We wish to determine the mechanistic basis for haspin action during mitosis. In Aim 1 we will use immunofluorescence and live cell imaging to examine the defects underlying chromosome misalignment caused by haspin depletion. We will define in detail defects in cohesion, chromosome-spindle attachment, spindle checkpoint activation, aurora B activity and spindle/centrosome function following haspin RNAi. In Aim 2 we will determine how H3 Thr-3 phosphorylation regulates centromeric chromatin. First, we will delineate the location of Thr-3 phosphorylation with respect to cohesin and other molecules, allowing refinement of current centromere structure models. Then we will use RNAi and overexpression to determine the influence of haspin on cohesin, chromosome passenger and heterochromatin protein binding at centromeres, and on patterns of histone modification. Expression of H3 molecules mutated at Thr-3 and in vitro binding studies will reveal the role of Thr-3 phosphorylation in these effects. In Aim 3 we explore functional interactions of haspin to understand its role in centrosome and spindle activity. Regulation of chromosome behavior during cell division is critical to allow accurate passage of the genome to daughter cells. Cancer cells have atypical numbers of abnormal chromosomes, suggesting that disruption of these mechanisms contributes to the generation of malignancy. A greater knowledge of the role of human haspin in this process will help us understand the defects that underlie transformation and may lead to new approaches to block the division of cancer cells.
Funding Period: ----------------2010 - ---------------2010-
more information: NIH RePORT
- Regulation of mitotic chromosome cohesion by Haspin and Aurora BJun Dai
Division of Rheumatology, Immunology, and Allergy, Department of Medicine, Brigham and Women s Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA
Dev Cell 11:741-50. 2006..We conclude that Haspin is required to maintain centromeric cohesion during mitosis. We also suggest that Aurora B regulates cohesin removal through its effect on the localization of Shugoshin...
- Haspin inhibitors reveal centromeric functions of Aurora B in chromosome segregationFangwei Wang
Division of Rheumatology, Immunology and Allergy, Brigham and Women s Hospital, Harvard Medical School, Boston, MA 02115, USA
J Cell Biol 199:251-68. 2012..Thus, Haspin inhibitors reveal centromeric roles of Aurora B in chromosome movement and spindle checkpoint signaling...
- Polo-like kinase-1 triggers histone phosphorylation by Haspin in mitosisLinli Zhou
Life Sciences Institute and Innovation Center for Cell Biology, Zhejiang University, Hangzhou, Zhejiang Province, China
EMBO Rep 15:273-81. 2014..These studies demonstrate that Plk1 can positively regulate CPC recruitment in mitosis. ..
- Phosphorylation of histone H3 at threonine 11 establishes a novel chromatin mark for transcriptional regulationEric Metzger
Universitäts Frauenklinik und Zentrale Klinische Forschung, Klinikum der Universität Freiburg, Breisacherstrasse 66, 79106 Freiburg, Germany
Nat Cell Biol 10:53-60. 2008..Finally, inhibition of PRK1 blocks proliferation of androgen receptor-induced tumour cell proliferation, making PRK1 a promising therapeutic target...
- Structure and functional characterization of the atypical human kinase haspinJeyanthy Eswaran
Structural Genomics Consortium, Nuffield Department of Medicine, University of Oxford, Oxford OX3 7DQ, United Kingdom
Proc Natl Acad Sci U S A 106:20198-203. 2009....
- Haspin: a newly discovered regulator of mitotic chromosome behaviorJonathan M G Higgins
Division of Rheumatology, Immunology and Allergy, Brigham and Women s Hospital, Harvard Medical School, 1 Jimmy Fund Way, Boston, MA 02115, USA
Chromosoma 119:137-47. 2010....
- Histone H3 Thr-3 phosphorylation by Haspin positions Aurora B at centromeres in mitosisFangwei Wang
Division of Rheumatology, Immunology, and Allergy, Brigham and Women s Hospital, Harvard Medical School, Smith Building, 1 Jimmy Fund Way, Boston, MA 02115, USA
Science 330:231-5. 2010..Therefore, H3T3ph generated by Haspin positions the CPC at centromeres to regulate selected targets of Aurora B during mitosis...
- Studies of haspin-depleted cells reveal that spindle-pole integrity in mitosis requires chromosome cohesionJun Dai
Brigham and Women s Hospital, Harvard Medical School, Boston, MA 02115, USA
J Cell Sci 122:4168-76. 2009....
- Perturbation of mitosis through inhibition of histone acetyltransferases: the key to ochratoxin a toxicity and carcinogenicity?Kristin Czakai
Department of Toxicology, Institute of Pharmacology and Toxicology, University of Wurzburg, Wurzburg, Germany
Toxicol Sci 122:317-29. 2011....
- A positive feedback loop involving Haspin and Aurora B promotes CPC accumulation at centromeres in mitosisFangwei Wang
Division of Rheumatology, Immunology and Allergy, Department of Medicine, Brigham and Women s Hospital, Harvard Medical School, Boston, MA 02115, USA
Curr Biol 21:1061-9. 2011..We also provide evidence that the Bub1-shugoshin-CPC pathway supplies a signal that boosts the CPC-Haspin-H3T3ph feedback loop specifically at centromeres to produce the well-known accumulation of the CPC in these regions...
- Molecular basis for phosphospecific recognition of histone H3 tails by Survivin paralogues at inner centromeresEwa Niedzialkowska
Department of Biochemistry and Molecular Genetics, University of Virginia School of Medicine, Charlottesville, VA 22908, USA
Mol Biol Cell 23:1457-66. 2012..Our data demonstrate that Survivin paralogues have different characteristics of phosphospecific binding to threonine-3 of histone H3, providing new insight into the biology of the inner centromere...
- A phospho/methyl switch at histone H3 regulates TFIID association with mitotic chromosomesRadhika A Varier
Department of Physiological Chemistry and Netherlands Proteomics Center, University Medical Centre Utrecht, Utrecht, The Netherlands
EMBO J 29:3967-78. 2010..Based on our observations, we propose that a histone H3 phospho-methyl switch regulates TFIID-mediated transcription during mitotic progression of the cell cycle...