Brief Communication | Published:

Light-induced nuclear export reveals rapid dynamics of epigenetic modifications

Nature Chemical Biology volume 12, pages 399401 (2016) | Download Citation

Abstract

We engineered a photoactivatable system for rapidly and reversibly exporting proteins from the nucleus by embedding a nuclear export signal in the LOV2 domain from phototropin 1. Fusing the chromatin modifier Bre1 to the photoswitch, we achieved light-dependent control of histone H2B monoubiquitylation in yeast, revealing fast turnover of the ubiquitin mark. Moreover, this inducible system allowed us to dynamically monitor the status of epigenetic modifications dependent on H2B ubiquitylation.

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Acknowledgements

We would like to thank D. Dickinson for help with C. elegans injections, H. Meriesh for providing a plasmid containing BRE1, and R. Dronamraju and Z.-W. Sun for helpful discussions. Funding: J.E.B. (NIH GM111557), B.K., K.H., B.S. (NIH DA036877), K.H. (GM102924).

Author information

Affiliations

  1. Department of Biochemistry and Biophysics, University of North Carolina, Chapel Hill, North Carolina, USA.

    • Hayretin Yumerefendi
    • , Andrew Michael Lerner
    • , Seth Parker Zimmerman
    • , Brian D Strahl
    •  & Brian Kuhlman
  2. Department of Pharmacology, University of North Carolina, Chapel Hill, North Carolina, USA.

    • Klaus Hahn
  3. Lineberger Comprehensive Cancer Center, University of North Carolina, Chapel Hill, North Carolina, USA.

    • Klaus Hahn
    • , James E Bear
    • , Brian D Strahl
    •  & Brian Kuhlman
  4. Department of Cell Biology & Physiology, University of North Carolina, Chapel Hill, North Carolina, USA.

    • James E Bear

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Contributions

H.Y. and B.K. conceived the project and designed the experiments. H.Y., A.M.L. and S.P.Z. performed the experiments. H.Y., A.M.L., S.P.Z., B.D.S. and B.K. analyzed the results. K.H. and J.E.B. provided reagents and equipment. H.Y. and B.K. wrote the manuscript with input from all authors.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Brian Kuhlman.

Supplementary information

PDF files

  1. 1.

    Supplementary Text and Figures

    Supplementary Results, Supplementary Figures 1–13, Supplementary Tables 1–4 and Supplementary Note.

Videos

  1. 1.

    LINXa3 blue light single activation and reversion in mouse fibroblast cells (IA32)

    The appearance and disappearance of a square indicate the area and the time of blue light activation. Scale bar is 50 μm.

  2. 2.

    LINXb3 blue light single activation and reversion in mouse fibroblast cells (IA32)

    The appearance and disappearance of a square indicate the area and the time of blue light activation. Scale bar is 50 μm.

  3. 3.

    Two cycles of blue light activation and reversion of LINXa3 in mouse fibroblast cells (IA32)

    The appearance and disappearance of a square indicate the area and the time of blue light activation. Scale bar is 100 μm.

  4. 4.

    Two cycles of blue light activation and reversion of LINXb3 in mouse fibroblast cells (IA32)

    The appearance and disappearance of a square indicate the area and the time of blue light activation. Scale bar is 50 μm.

  5. 5.

    Activation and reversion of LINXa3 in the C. elegans embryo

    Activation is performed on the entire field of view and is indicated with words at the upper left corner. Scale bar is 10 μm.

  6. 6.

    Two cycles of blue light activation and reversion of LINXa3-nano (red color) and iLID-Mito (green color) in mouse fibroblast cells (IA32)

    The appearance and disappearance of a square indicate the area and the time of blue light activation. Scale bar is 50 μm.

  7. 7.

    Activation and reversion of LINXa4-Bre1 (green color) in an H2B-mCherry (labels the nucleus in red) yeast strain (Supplementary Table 3)

    The appearance and disappearance of a square indicate the area and the time of blue light activation. Scale bar is 10 μm.

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DOI

https://doi.org/10.1038/nchembio.2068

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