Structure of the Lassa virus nucleoprotein revealed by X-ray crystallography, small-angle X-ray scattering, and electron microscopy.
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Structure of the Lassa virus nucleoprotein revealed by X-ray crystallography, small-angle X-ray scattering, and electron microscopy. / Brunotte, Linda; Kerber, Romy; Shang, Weifeng; Hauer, Florian; Hass, Meike; Gabriel, Martin; Lelke, Michaela; Busch, Carola; Stark, Holger; Svergun, Dmitri I; Betzel, Christian; Perbandt, Markus; Günther, Stephan.
in: J BIOL CHEM, Jahrgang 286, Nr. 44, 44, 2011, S. 38748-38756.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Structure of the Lassa virus nucleoprotein revealed by X-ray crystallography, small-angle X-ray scattering, and electron microscopy.
AU - Brunotte, Linda
AU - Kerber, Romy
AU - Shang, Weifeng
AU - Hauer, Florian
AU - Hass, Meike
AU - Gabriel, Martin
AU - Lelke, Michaela
AU - Busch, Carola
AU - Stark, Holger
AU - Svergun, Dmitri I
AU - Betzel, Christian
AU - Perbandt, Markus
AU - Günther, Stephan
PY - 2011
Y1 - 2011
N2 - The nucleoprotein (NP) of Lassa virus (LASV) strain AV was expressed in a recombinant baculovirus system. The crystal structure of full-length NP was solved at a resolution of 2.45 ?. The overall fold corresponds to that of NP of LASV strain Josiah (Qi, X., Lan, S., Wang, W., Schelde, L. M., Dong, H., Wallat, G. D., Ly, H., Liang, Y., and Dong, C. (2010) Nature 468, 779-783) with a root mean square deviation of 0.67 ? for all atoms (6.3% difference in primary sequence). As the packing in the crystal offers two different trimer architectures for the biological assembly, the quaternary structure of NP in solution was determined by small-angle x-ray scattering and EM. After classification and averaging of >6000 EM raw images, trimeric centrosymmetric structures were obtained, which correspond in size and shape to one trimer in the crystal structure formed around a crystallographic 3-fold rotation axis (symmetric trimer). The symmetric trimer is also a good model for the small-angle x-ray scattering data and could be well embedded into the ab initio model. The N-terminal domain of NP contains a deep nucleotide-binding cavity that has been proposed to bind cellular cap structures for priming viral mRNA synthesis. All residues implicated in m(7)GpppN binding were exchanged, and the transcription/replication phenotype of the NP mutant was tested using a LASV replicon system. None of the mutants showed a specific defect in mRNA expression; most were globally defective in RNA synthesis. In conclusion, we describe the full-length crystal structure and the quaternary structure in solution of LASV NP. The nucleotide-binding pocket of NP could not be assigned a specific role in viral mRNA synthesis.
AB - The nucleoprotein (NP) of Lassa virus (LASV) strain AV was expressed in a recombinant baculovirus system. The crystal structure of full-length NP was solved at a resolution of 2.45 ?. The overall fold corresponds to that of NP of LASV strain Josiah (Qi, X., Lan, S., Wang, W., Schelde, L. M., Dong, H., Wallat, G. D., Ly, H., Liang, Y., and Dong, C. (2010) Nature 468, 779-783) with a root mean square deviation of 0.67 ? for all atoms (6.3% difference in primary sequence). As the packing in the crystal offers two different trimer architectures for the biological assembly, the quaternary structure of NP in solution was determined by small-angle x-ray scattering and EM. After classification and averaging of >6000 EM raw images, trimeric centrosymmetric structures were obtained, which correspond in size and shape to one trimer in the crystal structure formed around a crystallographic 3-fold rotation axis (symmetric trimer). The symmetric trimer is also a good model for the small-angle x-ray scattering data and could be well embedded into the ab initio model. The N-terminal domain of NP contains a deep nucleotide-binding cavity that has been proposed to bind cellular cap structures for priming viral mRNA synthesis. All residues implicated in m(7)GpppN binding were exchanged, and the transcription/replication phenotype of the NP mutant was tested using a LASV replicon system. None of the mutants showed a specific defect in mRNA expression; most were globally defective in RNA synthesis. In conclusion, we describe the full-length crystal structure and the quaternary structure in solution of LASV NP. The nucleotide-binding pocket of NP could not be assigned a specific role in viral mRNA synthesis.
KW - Mutagenesis
KW - Protein Structure, Tertiary
KW - Transcription, Genetic
KW - Protein Binding
KW - Scattering, Radiation
KW - Molecular Conformation
KW - Mutation
KW - Crystallography, X-Ray/methods
KW - Protein Structure, Quaternary
KW - Lassa virus/chemistry/genetics/metabolism
KW - Microscopy, Electron/methods
KW - Nucleoproteins/chemistry/genetics
KW - RNA Viruses/chemistry
KW - X-Rays
KW - Mutagenesis
KW - Protein Structure, Tertiary
KW - Transcription, Genetic
KW - Protein Binding
KW - Scattering, Radiation
KW - Molecular Conformation
KW - Mutation
KW - Crystallography, X-Ray/methods
KW - Protein Structure, Quaternary
KW - Lassa virus/chemistry/genetics/metabolism
KW - Microscopy, Electron/methods
KW - Nucleoproteins/chemistry/genetics
KW - RNA Viruses/chemistry
KW - X-Rays
M3 - SCORING: Journal article
VL - 286
SP - 38748
EP - 38756
JO - J BIOL CHEM
JF - J BIOL CHEM
SN - 0021-9258
IS - 44
M1 - 44
ER -
