Large-Scale Conformational Dynamics Control H5N1 Influenza Polymerase PB2 Binding to Importin α
- Publication type:
- Journal article
- Metadata:
-
- Autoren
- Elise Delaforge
- Sigrid Milles
- Guillaume Bouvignies
- Denis Bouvier
- Stephane Boivin
- Nicola Salvi
- Damien Maurin
- Anne Martel
- Adam Round
- Edward A Lemke
- Malene Ringkjobing Jensen
- Darren J Hart
- Martin Blackledge
- Autoren-URL
- https://www.webofscience.com/api/gateway?GWVersion=2&SrcApp=fis-test-1&SrcAuth=WosAPI&KeyUT=WOS:000366339900014&DestLinkType=FullRecord&DestApp=WOS_CPL
- DOI
- 10.1021/jacs.5b07765
- eISSN
- 1520-5126
- Externe Identifier
- Clarivate Analytics Document Solution ID: CY3WF
- PubMed Identifier: 26424125
- ISSN
- 0002-7863
- Ausgabe der Veröffentlichung
- 48
- Zeitschrift
- JOURNAL OF THE AMERICAN CHEMICAL SOCIETY
- Paginierung
- 15122 - 15134
- Datum der Veröffentlichung
- 2015
- Status
- Published
- Titel
- Large-Scale Conformational Dynamics Control H5N1 Influenza Polymerase PB2 Binding to Importin α
- Sub types
- Article
- Ausgabe der Zeitschrift
- 137
Data source: Web of Science (Lite)
- Other metadata sources:
-
- Autoren
- Elise Delaforge
- Sigrid Milles
- Guillaume Bouvignies
- Denis Bouvier
- Stephane Boivin
- Nicola Salvi
- Damien Maurin
- Anne Martel
- Adam Round
- Edward A Lemke
- Malene Ringkjøbing Jensen
- Darren J Hart
- Martin Blackledge
- DOI
- 10.1021/jacs.5b07765
- eISSN
- 1520-5126
- ISSN
- 0002-7863
- Ausgabe der Veröffentlichung
- 48
- Zeitschrift
- Journal of the American Chemical Society
- Sprache
- en
- Online publication date
- 2015
- Paginierung
- 15122 - 15134
- Datum der Veröffentlichung
- 2015
- Status
- Published
- Herausgeber
- American Chemical Society (ACS)
- Herausgeber URL
- http://dx.doi.org/10.1021/jacs.5b07765
- Datum der Datenerfassung
- 2023
- Titel
- Large-Scale Conformational Dynamics Control H5N1 Influenza Polymerase PB2 Binding to Importin α
- Ausgabe der Zeitschrift
- 137
Data source: Crossref
- Abstract
- Influenza A RNA polymerase complex is formed from three components, PA, PB1, and PB2. PB2 is independently imported into the nucleus prior to polymerase reconstitution. All crystallographic structures of the PB2 C-terminus (residues 536-759) reveal two globular domains, 627 and NLS, that form a tightly packed heterodimer. The molecular basis of the affinity of 627-NLS for importins remained unclear from these structures, apparently requiring large-scale conformational changes prior to importin binding. Using a combination of solution-state NMR, small-angle neutron scattering, small-angle X-ray scattering (SAXS), and Förster resonance energy transfer (FRET), we show that 627-NLS populates a temperature-dependent dynamic equilibrium between closed and open states. The closed state is stabilized by a tripartite salt bridge involving the 627-NLS interface and the linker, that becomes flexible in the open state, with 627 and NLS dislocating into a highly dynamic ensemble. Activation enthalpies and entropies associated with the rupture of this interface were derived from simultaneous analysis of temperature-dependent chemical exchange saturation transfer measurements, revealing a strong temperature dependence of both open-state population and exchange rate. Single-molecule FRET and SAXS demonstrate that only the open-form is capable of binding to importin α and that, upon binding, the 627 domain samples a dynamic conformational equilibrium in the vicinity of the C-terminus of importin α. This intrinsic large-scale conformational flexibility therefore enables 627-NLS to bind importin through conformational selection from a temperature-dependent equilibrium comprising both functional forms of the protein.
- Addresses
- Univ. Grenoble Alpes , Institut de Biologie Structurale (IBS), F-38044 Grenoble, France.
- Autoren
- Elise Delaforge
- Sigrid Milles
- Guillaume Bouvignies
- Denis Bouvier
- Stephane Boivin
- Nicola Salvi
- Damien Maurin
- Anne Martel
- Adam Round
- Edward A Lemke
- Malene Ringkjøbing Jensen
- Darren J Hart
- Martin Blackledge
- DOI
- 10.1021/jacs.5b07765
- eISSN
- 1520-5126
- Externe Identifier
- PubMed Identifier: 26424125
- Funding acknowledgements
- Agence Nationale de la Recherche:
- R?gion Rh?ne -Alpes:
- Schweizerische Nationalfonds zur F?rderung der Wissenschaftlichen Forschung: P2ELP2_148858
- European Molecular Biology Organization:
- FINOVI Foundation:
- Open access
- false
- ISSN
- 0002-7863
- Ausgabe der Veröffentlichung
- 48
- Zeitschrift
- Journal of the American Chemical Society
- Schlüsselwörter
- Karyopherins
- Viral Proteins
- Solutions
- Crystallography, X-Ray
- Fluorescence Resonance Energy Transfer
- Nuclear Magnetic Resonance, Biomolecular
- Protein Conformation
- Protein Binding
- Influenza A Virus, H5N1 Subtype
- Sprache
- eng
- Medium
- Print-Electronic
- Online publication date
- 2015
- Paginierung
- 15122 - 15134
- Datum der Veröffentlichung
- 2015
- Status
- Published
- Datum der Datenerfassung
- 2015
- Titel
- Large-Scale Conformational Dynamics Control H5N1 Influenza Polymerase PB2 Binding to Importin α.
- Sub types
- Research Support, Non-U.S. Gov't
- Journal Article
- Ausgabe der Zeitschrift
- 137
Data source: Europe PubMed Central
- Abstract
- Influenza A RNA polymerase complex is formed from three components, PA, PB1, and PB2. PB2 is independently imported into the nucleus prior to polymerase reconstitution. All crystallographic structures of the PB2 C-terminus (residues 536-759) reveal two globular domains, 627 and NLS, that form a tightly packed heterodimer. The molecular basis of the affinity of 627-NLS for importins remained unclear from these structures, apparently requiring large-scale conformational changes prior to importin binding. Using a combination of solution-state NMR, small-angle neutron scattering, small-angle X-ray scattering (SAXS), and Förster resonance energy transfer (FRET), we show that 627-NLS populates a temperature-dependent dynamic equilibrium between closed and open states. The closed state is stabilized by a tripartite salt bridge involving the 627-NLS interface and the linker, that becomes flexible in the open state, with 627 and NLS dislocating into a highly dynamic ensemble. Activation enthalpies and entropies associated with the rupture of this interface were derived from simultaneous analysis of temperature-dependent chemical exchange saturation transfer measurements, revealing a strong temperature dependence of both open-state population and exchange rate. Single-molecule FRET and SAXS demonstrate that only the open-form is capable of binding to importin α and that, upon binding, the 627 domain samples a dynamic conformational equilibrium in the vicinity of the C-terminus of importin α. This intrinsic large-scale conformational flexibility therefore enables 627-NLS to bind importin through conformational selection from a temperature-dependent equilibrium comprising both functional forms of the protein.
- Autoren
- Elise Delaforge
- Sigrid Milles
- Guillaume Bouvignies
- Denis Bouvier
- Stephane Boivin
- Nicola Salvi
- Damien Maurin
- Anne Martel
- Adam Round
- Edward A Lemke
- Malene Ringkjøbing Jensen
- Darren J Hart
- Martin Blackledge
- Autoren-URL
- https://www.ncbi.nlm.nih.gov/pubmed/26424125
- DOI
- 10.1021/jacs.5b07765
- eISSN
- 1520-5126
- Ausgabe der Veröffentlichung
- 48
- Zeitschrift
- J Am Chem Soc
- Schlüsselwörter
- Crystallography, X-Ray
- Fluorescence Resonance Energy Transfer
- Influenza A Virus, H5N1 Subtype
- Karyopherins
- Nuclear Magnetic Resonance, Biomolecular
- Protein Binding
- Protein Conformation
- Solutions
- Viral Proteins
- Sprache
- eng
- Country
- United States
- Paginierung
- 15122 - 15134
- Datum der Veröffentlichung
- 2015
- Status
- Published
- Datum, an dem der Datensatz öffentlich gemacht wurde
- 2016
- Titel
- Large-Scale Conformational Dynamics Control H5N1 Influenza Polymerase PB2 Binding to Importin α.
- Sub types
- Journal Article
- Research Support, Non-U.S. Gov't
- Ausgabe der Zeitschrift
- 137
Data source: PubMed
- Beziehungen:
- Property of