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Structure of a mitochondrial fission dynamin in the closed conformation

Nature Structural & Molecular Biology volume 25, pages 722–731 (2018)Cite this article

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Abstract

Dynamin 1-like proteins (DNM1-L) are mechanochemical GTPases that induce membrane fission in mitochondria and peroxisomes. Their mechanism depends on conformational changes driven by nucleotide and lipid cycling. Here we show the crystal structure of a mitochondrial fission dynamin (CmDnm1) from the algae Cyanidioschyzon merolae. Unlike other eukaryotic dynamin structures, CmDnm1 is in a hinge 1 closed conformation, with the GTPase domain compacted against the stalk. Within the crystal, CmDnm1 packs as a diamond-shaped tetramer that is consistent with an inactive off-membrane state. Crosslinking, photoinduced electron transfer assays, and electron microscopy verify these structures. In vitro, CmDnm1 forms concentration-dependent rings and protein–lipid tubes reminiscent of DNM1-L and classical dynamin with hinge 1 open. Our data provides a mechanism for filament collapse and membrane release that may extend to other dynamin family members. Additionally, hinge 1 closing may represent a key conformational change that contributes to membrane fission.

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Fig. 1: CmDnm1 catalytic activity and self-assembly.
Fig. 2: The crystal structure of the CmDnm1 monomer.
Fig. 3: The crystal structure of the CmDnm1 tetramer.
Fig. 4: Biochemical validation of CmDnm1 hinge 1 closed conformation in solution.
Fig. 5: Validation of the CmDnm1 tetramer in solution.
Fig. 6: CmDnm1 nucleotide hydrolysis cycle and activation model.

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  • 08 August 2018

    In the version of this article initially published, the file with supplementary figures had formatting problems. This has now been corrected.

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Acknowledgements

We thank the Beamline staff at the ESRF for data collection support and T. Pape for in-house EM facility support. We acknowledge the gift of C. merolae genomic DNA from T. Kuroiwa. We are grateful to A. Chernyatina and J. Liu for manuscript feedback. This work was supported by a Wellcome Trust Fellowship (097328/Z/11/Z) to H.H.L. O.B. was supported by a BBSRC Doctoral Training Partnership with Imperial College.

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  1. Department of Life Sciences, Imperial College, London, UK

    Olga Bohuszewicz & Harry H. Low

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  1. Olga Bohuszewicz
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O.B. and H.H.L. designed experiments. H.H.L. initially purified native protein, obtained crystals, and determined structure. O.B. purified proteins and performed all other experiments including GTPase, crosslinking, and PET assays and generated EM data. H.H.L. and O.B. wrote the paper.

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Correspondence to Harry H. Low.

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Bohuszewicz, O., Low, H.H. Structure of a mitochondrial fission dynamin in the closed conformation. Nat Struct Mol Biol 25, 722–731 (2018). https://doi.org/10.1038/s41594-018-0097-6

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