Blog
About

11
views
0
recommends
+1 Recommend
0 collections
    0
    shares
      • Record: found
      • Abstract: found
      • Article: found
      Is Open Access

      The Structure of the R2TP Complex Defines a Platform for Recruiting Diverse Client Proteins to the HSP90 Molecular Chaperone System

      Read this article at

      Bookmark
          There is no author summary for this article yet. Authors can add summaries to their articles on ScienceOpen to make them more accessible to a non-specialist audience.

          Summary

          The R2TP complex, comprising the Rvb1p-Rvb2p AAA-ATPases, Tah1p, and Pih1p in yeast, is a specialized Hsp90 co-chaperone required for the assembly and maturation of multi-subunit complexes. These include the small nucleolar ribonucleoproteins, RNA polymerase II, and complexes containing phosphatidylinositol-3-kinase-like kinases. The structure and stoichiometry of yeast R2TP and how it couples to Hsp90 are currently unknown. Here, we determine the 3D organization of yeast R2TP using sedimentation velocity analysis and cryo-electron microscopy. The 359-kDa complex comprises one Rvb1p/Rvb2p hetero-hexamer with domains II (DIIs) forming an open basket that accommodates a single copy of Tah1p-Pih1p. Tah1p-Pih1p binding to multiple DII domains regulates Rvb1p/Rvb2p ATPase activity. Using domain dissection and cross-linking mass spectrometry, we identified a unique region of Pih1p that is essential for interaction with Rvb1p/Rvb2p. These data provide a structural basis for understanding how R2TP couples an Hsp90 dimer to a diverse set of client proteins and complexes.

          Graphical Abstract

          Highlights

          • Rvb1p-Rvb2p forms a hetero-hexamer with DII domains recruiting a single Tah1p-Pih1p
          • Residues 230–250 in Pih1p are essential to bind Rvb1p-Rvb2p
          • 3D structure of yeast R2TP couples an Hsp90 dimer to client proteins
          • Tah1p-Pih1p binding to flexible DII domains stimulates Rvb1p-Rvb2p ATPase activity

          Abstract

          Rivera-Calzada, Pal et al. reveal the 3D architecture of yeast R2TP using sedimentation velocity analysis, cryo-EM, XL/MS, and domain mapping, and find that the Rvb1p/Rvb2p ring binds a single Tah1p-Pih1p unit that modulates Rvb1/2-ATPase activity and couples phosphorylated clients and client adaptors to a single Hsp90 dimer.

          Related collections

          Most cited references 29

          • Record: found
          • Abstract: found
          • Article: found
          Is Open Access

          SWISS-MODEL: modelling protein tertiary and quaternary structure using evolutionary information

          Protein structure homology modelling has become a routine technique to generate 3D models for proteins when experimental structures are not available. Fully automated servers such as SWISS-MODEL with user-friendly web interfaces generate reliable models without the need for complex software packages or downloading large databases. Here, we describe the latest version of the SWISS-MODEL expert system for protein structure modelling. The SWISS-MODEL template library provides annotation of quaternary structure and essential ligands and co-factors to allow for building of complete structural models, including their oligomeric structure. The improved SWISS-MODEL pipeline makes extensive use of model quality estimation for selection of the most suitable templates and provides estimates of the expected accuracy of the resulting models. The accuracy of the models generated by SWISS-MODEL is continuously evaluated by the CAMEO system. The new web site allows users to interactively search for templates, cluster them by sequence similarity, structurally compare alternative templates and select the ones to be used for model building. In cases where multiple alternative template structures are available for a protein of interest, a user-guided template selection step allows building models in different functional states. SWISS-MODEL is available at http://swissmodel.expasy.org/.
            Bookmark
            • Record: found
            • Abstract: found
            • Article: not found

            Image processing for electron microscopy single-particle analysis using XMIPP.

            We describe a collection of standardized image processing protocols for electron microscopy single-particle analysis using the XMIPP software package. These protocols allow performing the entire processing workflow starting from digitized micrographs up to the final refinement and evaluation of 3D models. A particular emphasis has been placed on the treatment of structurally heterogeneous data through maximum-likelihood refinements and self-organizing maps as well as the generation of initial 3D models for such data sets through random conical tilt reconstruction methods. All protocols presented have been implemented as stand-alone, executable python scripts, for which a dedicated graphical user interface has been developed. Thereby, they may provide novice users with a convenient tool to quickly obtain useful results with minimum efforts in learning about the details of this comprehensive package. Examples of applications are presented for a negative stain random conical tilt data set on the hexameric helicase G40P and for a structurally heterogeneous data set on 70S Escherichia coli ribosomes embedded in vitrified ice.
              Bookmark
              • Record: found
              • Abstract: found
              • Article: not found

              Scipion: A software framework toward integration, reproducibility and validation in 3D electron microscopy.

              In the past few years, 3D electron microscopy (3DEM) has undergone a revolution in instrumentation and methodology. One of the central players in this wide-reaching change is the continuous development of image processing software. Here we present Scipion, a software framework for integrating several 3DEM software packages through a workflow-based approach. Scipion allows the execution of reusable, standardized, traceable and reproducible image-processing protocols. These protocols incorporate tools from different programs while providing full interoperability among them. Scipion is an open-source project that can be downloaded from http://scipion.cnb.csic.es.
                Bookmark

                Author and article information

                Contributors
                Journal
                Structure
                Structure
                Structure(London, England:1993)
                Cell Press
                0969-2126
                1878-4186
                05 July 2017
                05 July 2017
                : 25
                : 7
                : 1145-1152.e4
                Affiliations
                [1 ]Centro de Investigaciones Biológicas (CIB), Spanish National Research Council (CSIC), Ramiro de Maeztu 9, 28040 Madrid, Spain
                [2 ]Genome Damage and Stability Centre, School of Life Sciences, University of Sussex, Falmer, Brighton BN1 9RQ, UK
                [3 ]Structural Biology Unit, CIC bioGUNE, Bizkaia Technology Park, 48160 Derio, Spain
                [4 ]MRC Laboratory of Molecular Biology, Francis Crick Avenue, Cambridge CB2 0QH, UK
                [5 ]Structural Biology Programme, Spanish National Cancer Research Centre (CNIO), Melchor Fernández Almagro 3, 28029 Madrid, Spain
                Author notes
                []Corresponding author chris.prodromou@ 123456sussex.ac.uk
                [∗∗ ]Corresponding author laurence.pearl@ 123456sussex.ac.uk
                [∗∗∗ ]Corresponding author ollorca@ 123456cnio.es
                [6]

                These authors contributed equally

                [7]

                Lead Contact

                Article
                S0969-2126(17)30150-8
                10.1016/j.str.2017.05.016
                5501727
                28648606
                © 2017 The Authors

                This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).

                Categories
                Short Article

                Comments

                Comment on this article