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Optogenetic control of kinetochore function

Nature Chemical Biology volume 13, pages 1096–1101 (2017)Cite this article

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Abstract

Kinetochores act as hubs for multiple activities during cell division, including microtubule interactions and spindle checkpoint signaling. Each kinetochore can act autonomously, and activities change rapidly as proteins are recruited to, or removed from, kinetochores. Understanding this dynamic system requires tools that can manipulate kinetochores on biologically relevant temporal and spatial scales. Optogenetic approaches have the potential to provide temporal and spatial control with molecular specificity. Here we report new chemical inducers of protein dimerization that allow us to both recruit proteins to and release them from kinetochores using light. We use these dimerizers to manipulate checkpoint signaling and molecular motor activity. Our findings demonstrate specialized properties of the CENP-E (kinesin-7) motor for directional chromosome transport to the spindle equator and for maintenance of metaphase alignment. This work establishes a foundation for optogenetic control of kinetochore function, which is broadly applicable to experimental probing of other dynamic cellular processes.

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Figure 1: New dimerizers based on a modular design.
Figure 2: Activating and silencing the spindle checkpoint.
Figure 3: Kinesin-1 transports chromosomes in all directions from the pole.
Figure 4: CENP-E transports chromosomes from the pole to the equator.
Figure 5: CENP-E stabilizes metaphase alignment.

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Acknowledgements

We thank G. Furst and J. Gu for NMR assistance; R. Kohli for High-Resolution Mass Spectrometry (HRMS) assistance; A. Calderon and A. Gokden for assistance with molecular cloning and generating cell lines; D. Cleveland (University of California at San Diego) and E. Grishchuk (University of Pennsylvania) for CENP-E plasmids, L. Lavis for dye JF585 (Janelia Research Campus, HHMI); and E. Grishchuk and members of the Lampson lab for helpful discussions. C.A. thanks the Royal Thai Government for PhD fellowship funding through the Development and Promotion of Science and Technology (DPST) Project. This work was supported by the US National Institutes of Health (GM083988 to M.A.L. and GM118510 to D.M.C.) and the US National Institutes of Health, National Cancer Institute (U54-CA193417).

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Author notes

  1. Huaiying Zhang and Chanat Aonbangkhen: These authors contributed equally to this work.

Authors and Affiliations

  1. Department of Biology, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Huaiying Zhang, Ekaterina V Tarasovetc, Edward R Ballister & Michael A Lampson

  2. Department of Chemistry, School of Arts and Sciences, University of Pennsylvania, Philadelphia, Pennsylvania, USA

    Chanat Aonbangkhen & David M Chenoweth

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  1. Huaiying Zhang
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Contributions

H.Z. designed and conducted biological experiments and wrote the manuscript. C.A. synthesized and characterized the dimerizers, conducted experiments in Figure 1 and Supplementary Figures 1 and 2 and edited the manuscript. E.V.T. conducted experiments in Figure 4 and Supplementary Figure 4. E.R.B. contributed to the design of dimerizers and checkpoint experiments. D.M.C. and M.A.L. designed experiments and edited the manuscript.

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Correspondence to David M Chenoweth or Michael A Lampson.

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The authors declare no competing financial interests.

Supplementary information

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Supplementary Results and Supplementary Figures 1–4 (PDF 11468 kb)

Supplementary Note 1

Details of synthetic schemes and characterization, and supplementary references (PDF 3511 kb)

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Zhang, H., Aonbangkhen, C., Tarasovetc, E. et al. Optogenetic control of kinetochore function. Nat Chem Biol 13, 1096–1101 (2017). https://doi.org/10.1038/nchembio.2456

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