The Combination of X-Ray Crystallography and Cryo-Electron Microscopy Provides Insight into the Overall Architecture of the Dodecameric Rvb1/Rvb2 Complex

PLoS One. 2016 Jan 8;11(1):e0146457. doi: 10.1371/journal.pone.0146457. eCollection 2016.

Abstract

The Rvb1/Rvb2 complex is an essential component of many cellular pathways. The Rvb1/Rvb2 complex forms a dodecameric assembly where six copies of each subunit form two heterohexameric rings. However, due to conformational variability, the way the two rings pack together is still not fully understood. Here, we present the crystal structure and two cryo-electron microscopy reconstructions of the dodecameric, full-length Rvb1/Rvb2 complex, all showing that the interaction between the two heterohexameric rings is mediated through the Rvb1/Rvb2-specific domain II. Two conformations of the Rvb1/Rvb2 dodecamer are present in solution: a stretched conformation also present in the crystal, and a compact conformation. Novel asymmetric features observed in the reconstruction of the compact conformation provide additional insight into the plasticity of the Rvb1/Rvb2 complex.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Catalytic Domain
  • Chaetomium / enzymology*
  • Cryoelectron Microscopy
  • Crystallography, X-Ray
  • DNA Helicases / chemistry*
  • DNA-Binding Proteins / chemistry*
  • Fungal Proteins / chemistry*
  • Models, Molecular
  • Protein Interaction Domains and Motifs
  • Protein Structure, Quaternary
  • Protein Structure, Secondary

Substances

  • DNA-Binding Proteins
  • Fungal Proteins
  • DNA Helicases

Grants and funding

This work was funded by EMBL Interdisciplinary Postdoc programm under EU FP7 Marie-Curie Actions COFUND to NSM and CellNetworks (DFG-funded Excellence Initiative of the University of Heidelberg) to MID, PB, MB, CWM. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.