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Structural basis of oligomerization in the stalk region of dynamin-like MxA

Abstract

The interferon-inducible dynamin-like myxovirus resistance protein 1 (MxA; also called MX1) GTPase is a key mediator of cell-autonomous innate immunity against pathogens such as influenza viruses1. MxA partially localizes to COPI-positive membranes of the smooth endoplasmic reticulum–Golgi intermediate compartment2. At the point of infection, it redistributes to sites of viral replication and promotes missorting of essential viral constituents3,4. It has been proposed that the middle domain and the GTPase effector domain of dynamin-like GTPases constitute a stalk that mediates oligomerization and transmits conformational changes from the G domain to the target structure5,6,7; however, the molecular architecture of this stalk has remained elusive. Here we report the crystal structure of the stalk of human MxA, which folds into a four-helical bundle. This structure tightly oligomerizes in the crystal in a criss-cross pattern involving three distinct interfaces and one loop. Mutations in each of these interaction sites interfere with native assembly, oligomerization, membrane binding and antiviral activity of MxA. On the basis of these results, we propose a structural model for dynamin oligomerization and stimulated GTP hydrolysis that is consistent with previous structural predictions and has functional implications for all members of the dynamin family.

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Figure 1: Structure and oligomerization of the MxA stalk.
Figure 2: Biochemical analysis of the oligomerization interfaces.
Figure 3: Antiviral activity of MxA variants.
Figure 4: Oligomeric models of dynamin-like proteins.

Accession codes

Primary accessions

Protein Data Bank

Data deposits

The atomic coordinates of the MxA stalk have been deposited in the Protein Data Bank with accession number 3LJB.

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Acknowledgements

This project was supported by a grant of the Deutsche Forschungsgemeinschaft (SFB 740-From Molecules to Modules) and by a Career Development Fellowship of ‘The International Human Frontier Science Program Organization’ to O.D., and by the German-Israeli-Research foundation (GIF-841/04) to G.K. and O.H. We are grateful to J. Hinshaw and J. Mears for providing the EM maps and model fittings of oligomerized dynamin. We would also like to acknowledge help and support of O. Ristau and K. Schilling/Nanolytics (analytical ultracentrifugation), M. Dahte and H. Nikolenko (fluorescence measurements), A. Herrmann, T. Korte and P. Müller (stopped-flow analysis), G. Dittmar (mass spectrometry analysis), S. Werner and S. Gruber (technical assistance) and the BESSY staff at BL14.1 (data collection). This work was conducted by A.v.M. in partial fulfilment for a PhD degree from the Faculty of Biology at the University of Freiburg, Germany.

Author information

Authors and Affiliations

Authors

Contributions

S.G. solved the structure and carried out the biochemical characterization of MxA mutants. A.v.M. carried out all antiviral and cellular assays. S.P. assisted S.G. in cloning and purification. J.B. performed the analytical ultracentrifugation analysis. S.G., A.v.M., O.H., G.K. and O.D. planned the experimental design and wrote the manuscript.

Corresponding author

Correspondence to Oliver Daumke.

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The authors declare no competing financial interests.

Supplementary information

Supplementary Information

This file contains Supplementary Figure 1-12 with legends, Supplementary Tables 1-2 and References. (PDF 3284 kb)

Supplementary Data

This file contains the Pdb coordinates (dyn-oligomer.pdb) of four dynamin molecules in our proposed oligomer. The stalks of MxA were aligned as described in Fig. 4 and Supp. Fig. 12. The G-domains are from rat Dynamin1 (pdb 2AKA, ref. 8) with the position of the GDP molecule derived from Dictyostelium Dynamin (pdb 1JWY, ref. 61). The coordinates of the PH domain are from pdb 2DYN (ref. 62). (TXT 1400 kb)

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Gao, S., von der Malsburg, A., Paeschke, S. et al. Structural basis of oligomerization in the stalk region of dynamin-like MxA. Nature 465, 502–506 (2010). https://doi.org/10.1038/nature08972

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