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Title: Conformational landscape of a virus by single-particle X-ray scattering

Abstract

Using a manifold-based analysis of experimental diffraction snapshots from an X-ray free electron laser, we determine the three-dimensional structure and conformational landscape of the PR772 virus to a detector-limited resolution of 9 nm. Our results indicate that a single conformational coordinate controls reorganization of the genome, growth of a tubular structure from a portal vertex and release of the genome. Furthermore, these results demonstrate that single-particle X-ray scattering has the potential to shed light on key biological processes.

Authors:
 [1];  [1];  [1];  [1];  [1];  [1];  [1];  [1];  [2];  [3];  [4];  [2]; ORCiD logo [1]
  1. Univ. of Wisconsin, Milwaukee, WI (United States)
  2. SLAC National Accelerator Lab., Menlo Park, CA (United States)
  3. Arizona State Univ., Tempe, AZ (United States)
  4. Brookhaven National Lab. (BNL), Upton, NY (United States)
Publication Date:
Research Org.:
SLAC National Accelerator Lab., Menlo Park, CA (United States)
Sponsoring Org.:
USDOE
OSTI Identifier:
1394091
Grant/Contract Number:
AC02-76SF00515
Resource Type:
Journal Article: Accepted Manuscript
Journal Name:
Nature Methods
Additional Journal Information:
Journal Volume: 14; Journal Issue: 9; Journal ID: ISSN 1548-7091
Publisher:
Nature Publishing Group
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; 59 BASIC BIOLOGICAL SCIENCES; 97 MATHEMATICS AND COMPUTING

Citation Formats

Hosseinizadeh, Ahmad, Mashayekhi, Ghoncheh, Copperman, Jeremy, Schwander, Peter, Dashti, Ali, Sepehr, Reyhaneh, Fung, Russell, Schmidt, Marius, Yoon, Chun Hong, Hogue, Brenda G., Williams, Garth J., Aquila, Andrew, and Ourmazd, Abbas. Conformational landscape of a virus by single-particle X-ray scattering. United States: N. p., 2017. Web. doi:10.1038/NMETH.4395.
Hosseinizadeh, Ahmad, Mashayekhi, Ghoncheh, Copperman, Jeremy, Schwander, Peter, Dashti, Ali, Sepehr, Reyhaneh, Fung, Russell, Schmidt, Marius, Yoon, Chun Hong, Hogue, Brenda G., Williams, Garth J., Aquila, Andrew, & Ourmazd, Abbas. Conformational landscape of a virus by single-particle X-ray scattering. United States. doi:10.1038/NMETH.4395.
Hosseinizadeh, Ahmad, Mashayekhi, Ghoncheh, Copperman, Jeremy, Schwander, Peter, Dashti, Ali, Sepehr, Reyhaneh, Fung, Russell, Schmidt, Marius, Yoon, Chun Hong, Hogue, Brenda G., Williams, Garth J., Aquila, Andrew, and Ourmazd, Abbas. 2017. "Conformational landscape of a virus by single-particle X-ray scattering". United States. doi:10.1038/NMETH.4395.
@article{osti_1394091,
title = {Conformational landscape of a virus by single-particle X-ray scattering},
author = {Hosseinizadeh, Ahmad and Mashayekhi, Ghoncheh and Copperman, Jeremy and Schwander, Peter and Dashti, Ali and Sepehr, Reyhaneh and Fung, Russell and Schmidt, Marius and Yoon, Chun Hong and Hogue, Brenda G. and Williams, Garth J. and Aquila, Andrew and Ourmazd, Abbas},
abstractNote = {Using a manifold-based analysis of experimental diffraction snapshots from an X-ray free electron laser, we determine the three-dimensional structure and conformational landscape of the PR772 virus to a detector-limited resolution of 9 nm. Our results indicate that a single conformational coordinate controls reorganization of the genome, growth of a tubular structure from a portal vertex and release of the genome. Furthermore, these results demonstrate that single-particle X-ray scattering has the potential to shed light on key biological processes.},
doi = {10.1038/NMETH.4395},
journal = {Nature Methods},
number = 9,
volume = 14,
place = {United States},
year = 2017,
month = 8
}

Journal Article:
Free Publicly Available Full Text
This content will become publicly available on August 14, 2018
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  • Alphaviruses, such as Sindbis virus, undergo dramatic changes in three-dimensional structure upon exposure to low pH, and such exposure can establish conditions allowing fusion of the virus membrane with a cell plasma membrane upon return to neutral pH. While exposure to low pH is not required for entry of Sindbis virus into vertebrate or invertebrate cells, the conformational changes occurring at low pH may mimic those occurring upon virus-receptor interaction. Here, we employed small-angle neutron scattering with contrast variation to probe how the structure of a mammalian-grown Sindbis virus responds to moderately acidic pH. Several changes took place throughout themore » virion structure when the pH decreased from 7.2 to 6.4. Specifically, the RNA in the virion core underwent a conformational change. Additionally, the protein was redistributed. A significant amount of protein moved from the layer containing the lipid bilayer to the exterior of the virion. The results improve our understanding of the pH-driven alteration of Sindbis virus structure.« less
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  • Determining the interconverting conformations of dynamic proteins in atomic detail is a major challenge for structural biology. Conformational heterogeneity in the active site of the dynamic enzyme cyclophilin A (CypA) has been previously linked to its catalytic function, but the extent to which the different conformations of these residues are correlated is unclear. Here we compare the conformational ensembles of CypA by multitemperature synchrotron crystallography and fixed-target X-ray free-electron laser (XFEL) crystallography. The diffraction-before-destruction nature of XFEL experiments provides a radiation-damage-free view of the functionally important alternative conformations of CypA, confirming earlier synchrotron-based results. We monitored the temperature dependences ofmore » these alternative conformations with eight synchrotron datasets spanning 100-310 K. Multiconformer models show that many alternative conformations in CypA are populated only at 240 K and above, yet others remain populated or become populated at 180 K and below. These results point to a complex evolution of conformational heterogeneity between 180-–240 K that involves both thermal deactivation and solvent-driven arrest of protein motions in the crystal. The lack of a single shared conformational response to temperature within the dynamic active-site network provides evidence for a conformation shuffling model, in which exchange between rotamer states of a large aromatic ring in the middle of the network shifts the conformational ensemble for the other residues in the network. Together, our multitemperature analyses and XFEL data motivate a new generation of temperature- and time-resolved experiments to structurally characterize the dynamic underpinnings of protein function.« less
    Cited by 14
  • Determining the interconverting conformations of dynamic proteins in atomic detail is a major challenge for structural biology. Conformational heterogeneity in the active site of the dynamic enzyme cyclophilin A (CypA) has been previously linked to its catalytic function, but the extent to which the different conformations of these residues are correlated is unclear. Here we compare the conformational ensembles of CypA by multitemperature synchrotron crystallography and fixed-target X-ray free-electron laser (XFEL) crystallography. The diffraction-before-destruction nature of XFEL experiments provides a radiation-damage-free view of the functionally important alternative conformations of CypA, confirming earlier synchrotron-based results. We monitored the temperature dependences ofmore » these alternative conformations with eight synchrotron datasets spanning 100-310 K. Multiconformer models show that many alternative conformations in CypA are populated only at 240 K and above, yet others remain populated or become populated at 180 K and below. These results point to a complex evolution of conformational heterogeneity between 180-–240 K that involves both thermal deactivation and solvent-driven arrest of protein motions in the crystal. The lack of a single shared conformational response to temperature within the dynamic active-site network provides evidence for a conformation shuffling model, in which exchange between rotamer states of a large aromatic ring in the middle of the network shifts the conformational ensemble for the other residues in the network. Altogether, our multitemperature analyses and XFEL data motivate a new generation of temperature- and time-resolved experiments to structurally characterize the dynamic underpinnings of protein function.« less