TY  - JOUR
AU  - Feuerstein, S.
AU  - Solyom, Z.
AU  - Aladag, A.
AU  - Favier, A.
AU  - Schwarten, M.
AU  - Hoffmann, S.
AU  - Willbold, D.
AU  - Brutscher, B.
TI  - Transient structure and SH3 interaction sites in an intrinsically disordered fragment of the hepatitis C virus protein NS5A.
JO  - Journal of molecular biology
VL  - 420
IS  - 4-5
SN  - 0022-2836
CY  - Amsterdam [u.a.]
PB  - Elsevier
M1  - PreJuSER-21695
SP  - 310–323
PY  - 2012
N1  - 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).
AB  - 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.
KW  - Adaptor Proteins, Signal Transducing: metabolism
KW  - Binding Sites
KW  - Electron Spin Resonance Spectroscopy
KW  - Humans
KW  - Magnetic Resonance Spectroscopy
KW  - Nuclear Proteins: metabolism
KW  - Proline: chemistry
KW  - Protein Binding
KW  - Protein Structure, Tertiary
KW  - Scattering, Small Angle
KW  - Tumor Suppressor Proteins: metabolism
KW  - Viral Nonstructural Proteins: chemistry
KW  - Viral Nonstructural Proteins: genetics
KW  - Viral Nonstructural Proteins: metabolism
KW  - X-Rays
KW  - src Homology Domains
KW  - Adaptor Proteins, Signal Transducing (NLM Chemicals)
KW  - BIN1 protein, human (NLM Chemicals)
KW  - NS-5 protein, hepatitis C virus (NLM Chemicals)
KW  - Nuclear Proteins (NLM Chemicals)
KW  - Tumor Suppressor Proteins (NLM Chemicals)
KW  - Viral Nonstructural Proteins (NLM Chemicals)
KW  - Proline (NLM Chemicals)
KW  - J (WoSType)
LB  - PUB:(DE-HGF)16
C6  - pmid:22543239
UR  - <Go to ISI:>//WOS:000306250400005
DO  - DOI:10.1016/j.jmb.2012.04.023
UR  - https://juser.fz-juelich.de/record/21695
ER  -