Journal Article

PreJuSER-21695

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Transient structure and SH3 interaction sites in an intrinsically disordered fragment of the hepatitis C virus protein NS5A.

Feuerstein, S.FZJ* ; Solyom, Z.FZJ* ; Aladag, A.FZJ* ; Favier, A.FZJ* ; Schwarten, M.FZJ* ; Hoffmann, S.FZJ* ; Willbold, D.FZJ* ; Brutscher, B.FZJ*

2012
Elsevier Amsterdam [u.a.]

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Please use a persistent id in citations: doi:10.1016/j.jmb.2012.04.023

Abstract: Understanding the molecular mechanisms involved in virus replication and particle assembly is of primary fundamental and biomedical importance. Intrinsic conformational disorder plays a prominent role in viral proteins and their interaction with other viral and host cell proteins via transiently populated structural elements. Here, we report on the results of an investigation of an intrinsically disordered 188-residue fragment of the hepatitis C virus non-structural protein 5A (NS5A), which contains a classical poly-proline Src homology 3 (SH3) binding motif, using sensitivity- and resolution-optimized multidimensional NMR methods, complemented by small-angle X-ray scattering data. Our study provides detailed atomic-resolution information on transient local and long-range structure, as well as fast time scale dynamics in this NS5A fragment. In addition, we could characterize two distinct interaction modes with the SH3 domain of Bin1 (bridging integrator protein 1), a pro-apoptotic tumor suppressor. Despite being largely disordered, the protein contains three regions that transiently adopt α-helical structures, partly stabilized by long-range tertiary interactions. Two of these transient α-helices form a noncanonical SH3-binding motif, which allows low-affinity SH3 binding. Our results contribute to a better understanding of the role of the NS5A protein during hepatitis C virus infection. The present work also highlights the power of NMR spectroscopy to characterize multiple binding events including short-lived transient interactions between globular and highly disordered proteins.

Keyword(s): Adaptor Proteins, Signal Transducing: metabolism (MeSH) ; Binding Sites (MeSH) ; Electron Spin Resonance Spectroscopy (MeSH) ; Humans (MeSH) ; Magnetic Resonance Spectroscopy (MeSH) ; Nuclear Proteins: metabolism (MeSH) ; Proline: chemistry (MeSH) ; Protein Binding (MeSH) ; Protein Structure, Tertiary (MeSH) ; Scattering, Small Angle (MeSH) ; Tumor Suppressor Proteins: metabolism (MeSH) ; Viral Nonstructural Proteins: chemistry (MeSH) ; Viral Nonstructural Proteins: genetics (MeSH) ; Viral Nonstructural Proteins: metabolism (MeSH) ; X-Rays (MeSH) ; src Homology Domains (MeSH) ; Adaptor Proteins, Signal Transducing ; BIN1 protein, human ; NS-5 protein, hepatitis C virus ; Nuclear Proteins ; Tumor Suppressor Proteins ; Viral Nonstructural Proteins ; Proline ; J ; Bin1 (auto) ; hepatitis C virus (auto) ; intrinsic disorder (auto) ; NMR (auto) ; NS5A (auto)

Note: The authors are grateful to Isabel Ayala, Ombeline Pessey, and Lionel Imbert for help in protein production. We also acknowledge access to the EMBL X33 beamline at the DORIS storage ring, Deutsches Elektronen-Synchrotron, Hamburg, and we thank Clement Blanchet, Giancarlo Tria, and Dmitri Svergun for technical support and assistance with data analysis. This work was supported by the Commisariat a lEnergie Atomique et aux Energies Alternatives, the Centre National de la Recherche Scientifique, the University Grenoble1, and the Deutsche Forschungsgemeinschaft (SFB575), and by a European Marie-Curie grant (FP7-ITN-TDP-byNMR contract No. 264257).

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Contributing Institute(s):

1. Strukturbiochemie (ICS-6)

Research Program(s):

1. Funktion und Dysfunktion des Nervensystems (P33)
2. BioSoft: Makromolekulare Systeme und biologische Informationsverarbeitung (P45)
3. IDPBYNMR - High resolution tools to understand the functional role of protein intrinsic disorder (264257) (264257)

Appears in the scientific report 2012

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Database coverage:
; BIOSIS Previews ; Current Contents - Life Sciences ; JCR ; NCBI Molecular Biology Database ; Nationallizenz ; SCOPUS ; Science Citation Index ; Science Citation Index Expanded ; Thomson Reuters Master Journal List ; Web of Science Core Collection

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