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H2B Tyr37 phosphorylation suppresses expression of replication-dependent core histone genes

Nature Structural & Molecular Biology volume 19, pages 930–937 (2012)Cite this article

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Abstract

Histone gene transcription is actively downregulated after completion of DNA synthesis to avoid overproduction. However, the precise mechanistic details of the cessation of histone mRNA synthesis are not clear. We found that histone H2B phosphorylation at Tyr37 occurs upstream of histone cluster 1, Hist1, during the late S phase. We identified WEE1 as the kinase that phosphorylates H2B at Tyr37. Loss of expression or inhibition of WEE1 kinase abrogated H2B Tyr37 phosphorylation with a concomitant increase in histone transcription in yeast and mammalian cells. H2B Tyr37 phosphorylation excluded binding of the transcriptional coactivator NPAT and RNA polymerase II and recruited the histone chaperone HIRA upstream of the Hist1 cluster. Taken together, our data show a previously unknown and evolutionarily conserved function for WEE1 kinase as an epigenetic modulator that marks chromatin with H2B Tyr37 phosphorylation, thereby inhibiting the transcription of multiple histone genes to lower the burden on the histone mRNA turnover machinery.

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Figure 1: WEE1 phosphorylates histone H2B at Tyr37.
Figure 2: H2BpY37 occurs upstream of the histone gene cluster Hist1.
Figure 3: H2BpY37 suppresses the transcription of histone genes located in the Hist1 cluster.
Figure 4: Histone tyrosine phosphorylation by SWE1 is required for the transcriptional suppression of histones in yeast.
Figure 5: H2BpY37 differentially regulates NPAT and HIRA recruitment.
Figure 6: A model of H2B Tyr37 phosphorylation upstream of the major histone gene cluster Hist1 suppresses histone mRNA transcription.

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Acknowledgements

We thank A. Shilatifard (Stowers Institute) for WT and the Y40A mutant of S. cerevisiae, M. Mollapour (US National Institutes of Health) for WT and the swe1Δ mutant of S. cerevisiae and G. Enders (Fox Chase Cancer Center) for myc-tagged WEE1 constructs. We thank X. Qu for bioinformatics analysis; L. Hall and J. Repass for qRT-PCR analysis; K. Shapland and J. Kroeger for flow cytometry; and E. Seto for critical reading of the manuscript. We thank the Moffitt Flow Cytometry, Molecular Biology core facilities, Lung Cancer Spore and the Comprehensive Melanoma Research Center. The project was supported by a Moffitt Support grant to N.P.M.

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Authors and Affiliations

  1. Drug Discovery, Moffitt Cancer Center, Tampa, Florida, USA

    Kiran Mahajan & Nupam P Mahajan

  2. Proteomics Facility, Moffitt Cancer Center, Tampa, Florida, USA

    Bin Fang & John M Koomen

  3. Department of Oncologic Sciences, University of South Florida, Tampa, Florida, USA

    Nupam P Mahajan

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Contributions

K.M. and N.P.M. conceived the idea and designed all the experiments. K.M. and N.P.M. performed all the experiments except the mass spectrometric identification of H2B Tyr37 phosphorylation, which was performed by B.F. and J.M.K. K.M. and N.P.M. analyzed data and wrote the manuscript.

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Correspondence to Nupam P Mahajan.

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K.M. and N.P.M. are named as inventors on US patent application 61/583,864 titled “Antibodies specific for phosphorylated histones and uses thereof.”

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Mahajan, K., Fang, B., Koomen, J. et al. H2B Tyr37 phosphorylation suppresses expression of replication-dependent core histone genes. Nat Struct Mol Biol 19, 930–937 (2012). https://doi.org/10.1038/nsmb.2356

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