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Cryo-EM reveals different coronin binding modes for ADP– and ADP–BeFx actin filaments

Nature Structural & Molecular Biology volume 21, pages 10751081 (2014) | Download Citation

Abstract

Essential cellular processes involving the actin cytoskeleton are regulated by auxiliary proteins that can sense the nucleotide state of actin. Here we report cryo-EM structures for ADP-bound and ADP–beryllium fluoride (ADP–BeFx, an ADP-Pi mimic)-bound actin filaments in complex with the β-propeller domain of yeast coronin 1 (crn1), at 8.6-Å resolution. Our structures reveal the main differences in the interaction of coronin with the two nucleotide states of F-actin. We derived pseudoatomic models by fitting the atomic structures of actin and coronin into the EM envelopes and confirmed the identified interfaces on actin by chemical cross-linking, fluorescence spectroscopy and actin mutagenesis. The models offer a structural explanation for the nucleotide-dependent effects of coronin on cofilin-assisted remodeling of F-actin.

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Acknowledgements

This work was supported by US National Institutes of Health (US NIH) grants GM077190 (to E.R.), GM071940 and AI094386 (to Z.H.Z.) and F32HL119069 (to Z.A.O.D.); an American Heart Association postdoctoral fellowship 13POST17340020 (to P.G.); and a startup fund from The Ohio State University (to D.S.K.). The authors acknowledge the use of instruments at the Electron Imaging Center for NanoMachines supported by US NIH grant 1S10RR23057 (to Z.H.Z.) and CNSI at UCLA. The authors also acknowledge the use of computer time at the Extreme Science and Engineering Discovery Environment (XSEDE) resources (MCB130126 to Z.H.Z.) and thank B. Goode (Brandeis University) for coronin-expression plasmids. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health.

Author information

Author notes

    • Peng Ge
    •  & Zeynep A Oztug Durer

    These authors contributed equally to this work.

Affiliations

  1. Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles (UCLA), Los Angeles, California, USA.

    • Peng Ge
    •  & Z Hong Zhou
  2. California NanoSystems Institute (CNSI), UCLA, Los Angeles, California, USA.

    • Peng Ge
    •  & Z Hong Zhou
  3. Department of Chemistry and Biochemistry, UCLA, Los Angeles, California, USA.

    • Zeynep A Oztug Durer
    •  & Emil Reisler
  4. Department of Chemistry and Biochemistry, Ohio State University, Columbus, Ohio, USA.

    • Dmitri Kudryashov
  5. Molecular Biology Institute, UCLA, Los Angeles, California, USA.

    • Z Hong Zhou
    •  & Emil Reisler

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Contributions

P.G., Z.A.O.D., D.K. and E.R. designed experiments; P.G., Z.A.O.D. and D.K., collected EM data; P.G. and Z.H.Z. processed, analyzed and interpreted EM data; Z.A.O.D. collected and analyzed biochemical data; all authors wrote and reviewed the manuscript.

Competing interests

The authors declare no competing financial interests.

Corresponding author

Correspondence to Emil Reisler.

Integrated supplementary information

Supplementary information

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  1. 1.

    Supplementary Text and Figures

    Supplementary Figures 1–6 and Supplementary Table 1

Videos

  1. 1.

    Comparison between the pseudoatomic models of F-actin–coronin complexes in ADP and ADP–BeFx states.

    The two pseudo-atomic models are morphed back and forth as a movie in two orthogonal views. The movie starts with the viewpoint similar to Figure 2a, showing the model in ADP state. It then morphs to ADP-BeFx state and back to ADP state. The model then rotates 90° about its helical axis, and performs the same morphing again.

  2. 2.

    Animation of the competition between coronin and cofilin in ADP–BeFx state, not in ADP state (based on Figure 6).

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DOI

https://doi.org/10.1038/nsmb.2907

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