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Conformational changes of the multifunction p97 AAA ATPase during its ATPase cycle

Nature Structural Biology volume 9, pages 950–957 (2002)Cite this article

Abstract

p97 (also called VCP), a member of the AAA ATPase family, is involved in several cellular processes, including membrane fusion and extraction of proteins from the endoplasmic reticulum for cytoplasmic degradation. We have studied the conformational changes that p97 undergoes during the ATPase cycle by cryo-EM and single-particle analysis. Three-dimensional maps show that the two AAA domains, D1 and D2, as well as the N-domains, experience conformational changes during ATP binding, ATP hydrolysis, Pi release and ADP release. The N-domain is flexible in most nucleotide states except after ATP hydrolysis. The rings formed by D1 and D2 rotate with respect to each other, and the size of their axial openings fluctuates. Taken together, our results depict the movements that this and possibly other AAA ATPases can undergo during an ATPase cycle.

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Figure 1: p97 and ΔN-p97.
Figure 2: Structure of p97 in different nucleotide conditions.
Figure 3: Conformational changes of p97.
Figure 4: Stucture of ΔN-p97.

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Acknowledgements

This work was supported by National Institutes of Health grants to R.A.M., E.M.W.-K. and W.I.W. A.P.M. was funded by a Wellcome Trust Prize International Traveling Fellowship. We thank P. Chacon for doing the fitting with Situs47 and B. Sheehan for computing help. We also thank S. Kaiser, M. Bowen and C. Moores for critical reading of the manuscript.

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Author notes

  1. Isabelle Rouiller

    Present address: The Burnham Institute, 10901 North Torrey Pines Road, La Jolla, California, 92037, USA

  2. Andrew P. May

    Present address: Signature Bioscience, 475 Brannan Street, San Francisco, California, 94107, USA

Authors and Affiliations

  1. The Department of Cell Biology, The Scripps Research Institute, 10550 North Torrey Pines Road, La Jolla, 92037, California, USA

    Isabelle Rouiller, Ronald A. Milligan & Elizabeth M. Wilson-Kubalek

  2. Howard Hughes Medical Institute and Departments of Molecular and Cellular Physiology, Neurology and Neurological Sciences, and Stanford Synchrotron Radiation Laboratory, Stanford University, Stanford, 94305-5489, California, USA

    Byron DeLaBarre & Axel T. Brunger

  3. Departments of Structural Biology and Molecular and Cellular Physiology, Stanford University, Stanford, 94305-5126, California, USA

    Andrew P. May & William I. Weis

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Correspondence to Elizabeth M. Wilson-Kubalek.

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Rouiller, I., DeLaBarre, B., May, A. et al. Conformational changes of the multifunction p97 AAA ATPase during its ATPase cycle. Nat Struct Mol Biol 9, 950–957 (2002). https://doi.org/10.1038/nsb872

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