Coronavirus N protein N-terminal domain (NTD) specifically binds the transcriptional regulatory sequence (TRS) and melts TRS-cTRS RNA duplexes.
Identifieur interne : 002B53 ( Main/Exploration ); précédent : 002B52; suivant : 002B54Coronavirus N protein N-terminal domain (NTD) specifically binds the transcriptional regulatory sequence (TRS) and melts TRS-cTRS RNA duplexes.
Auteurs : Nicholas E. Grossoehme [États-Unis] ; Lichun Li ; Sarah C. Keane ; Pinghua Liu ; Charles E. Dann ; Julian L. Leibowitz ; David P. GiedrocSource :
- Journal of molecular biology [ 1089-8638 ] ; 2009.
Descripteurs français
- KwdFr :
- ARN viral (), ARN viral (génétique), ARN viral (métabolisme), Animaux, Cinétique, Conformation d'acide nucléique, Cristallographie aux rayons X, Données de séquences moléculaires, Liaison aux protéines, Modèles moléculaires, Mutagenèse dirigée, Protéines nucléocapside (), Protéines nucléocapside (génétique), Protéines nucléocapside (métabolisme), Protéines recombinantes (), Protéines recombinantes (génétique), Protéines recombinantes (métabolisme), Similitude de séquences d'acides aminés, Souris, Structure tertiaire des protéines, Séquence d'acides aminés, Séquence nucléotidique, Thermodynamique, Virus de l'hépatite murine (génétique), Virus de l'hépatite murine (métabolisme), Virus du SRAS (génétique), Virus du SRAS (métabolisme), Électricité statique, Éléments de régulation transcriptionnelle.
- MESH :
- génétique : ARN viral, Protéines nucléocapside, Protéines recombinantes, Virus de l'hépatite murine, Virus du SRAS.
- métabolisme : ARN viral, Protéines nucléocapside, Protéines recombinantes, Virus de l'hépatite murine, Virus du SRAS.
- ARN viral, Animaux, Cinétique, Conformation d'acide nucléique, Cristallographie aux rayons X, Données de séquences moléculaires, Liaison aux protéines, Modèles moléculaires, Mutagenèse dirigée, Protéines nucléocapside, Protéines recombinantes, Similitude de séquences d'acides aminés, Souris, Structure tertiaire des protéines, Séquence d'acides aminés, Séquence nucléotidique, Thermodynamique, Électricité statique, Éléments de régulation transcriptionnelle.
English descriptors
- KwdEn :
- Amino Acid Sequence, Animals, Base Sequence, Crystallography, X-Ray, Kinetics, Mice, Models, Molecular, Molecular Sequence Data, Murine hepatitis virus (genetics), Murine hepatitis virus (metabolism), Mutagenesis, Site-Directed, Nucleic Acid Conformation, Nucleocapsid Proteins (chemistry), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (metabolism), Protein Binding, Protein Structure, Tertiary, RNA, Viral (chemistry), RNA, Viral (genetics), RNA, Viral (metabolism), Recombinant Proteins (chemistry), Recombinant Proteins (genetics), Recombinant Proteins (metabolism), Regulatory Elements, Transcriptional, SARS Virus (genetics), SARS Virus (metabolism), Sequence Homology, Amino Acid, Static Electricity, Thermodynamics.
- MESH :
- chemical , chemistry : Nucleocapsid Proteins, RNA, Viral, Recombinant Proteins.
- genetics : Murine hepatitis virus, Nucleocapsid Proteins, RNA, Viral, Recombinant Proteins, SARS Virus.
- metabolism : Murine hepatitis virus, Nucleocapsid Proteins, RNA, Viral, Recombinant Proteins, SARS Virus.
- Amino Acid Sequence, Animals, Base Sequence, Crystallography, X-Ray, Kinetics, Mice, Models, Molecular, Molecular Sequence Data, Mutagenesis, Site-Directed, Nucleic Acid Conformation, Protein Binding, Protein Structure, Tertiary, Regulatory Elements, Transcriptional, Sequence Homology, Amino Acid, Static Electricity, Thermodynamics.
Abstract
All coronaviruses (CoVs), including the causative agent of severe acute respiratory syndrome (SARS), encode a nucleocapsid (N) protein that harbors two independent RNA binding domains of known structure, but poorly characterized RNA binding properties. We show here that the N-terminal domain (NTD) of N protein from mouse hepatitis virus (MHV), a virus most closely related to SARS-CoV, employs aromatic amino acid-nucleobase stacking interactions with a triple adenosine motif to mediate high-affinity binding to single-stranded RNAs containing the transcriptional regulatory sequence (TRS) or its complement (cTRS). Stoichiometric NTD fully unwinds a TRS-cTRS duplex that mimics a transiently formed transcription intermediate in viral subgenomic RNA synthesis. Mutation of the solvent-exposed Y127, positioned on the beta-platform surface of our 1.75 A structure, binds the TRS far less tightly and is severely crippled in its RNA unwinding activity. In contrast, the C-terminal domain (CTD) exhibits no RNA unwinding activity. Viruses harboring Y127A N mutation are strongly selected against and Y127A N does not support an accessory function in MHV replication. We propose that the helix melting activity of the coronavirus N protein NTD plays a critical accessory role in subgenomic RNA synthesis and other processes requiring RNA remodeling.
DOI: 10.1016/j.jmb.2009.09.040
PubMed: 19782089
Affiliations:
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Le document en format XML
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(metabolism)</term><term>Recombinant Proteins (chemistry)</term><term>Recombinant Proteins (genetics)</term><term>Recombinant Proteins (metabolism)</term><term>Regulatory Elements, Transcriptional</term><term>SARS Virus (genetics)</term><term>SARS Virus (metabolism)</term><term>Sequence Homology, Amino Acid</term><term>Static Electricity</term><term>Thermodynamics</term></keywords><keywords scheme="KwdFr" xml:lang="fr"><term>ARN viral ()</term><term>ARN viral (génétique)</term><term>ARN viral (métabolisme)</term><term>Animaux</term><term>Cinétique</term><term>Conformation d'acide nucléique</term><term>Cristallographie aux rayons X</term><term>Données de séquences moléculaires</term><term>Liaison aux protéines</term><term>Modèles moléculaires</term><term>Mutagenèse dirigée</term><term>Protéines nucléocapside ()</term><term>Protéines nucléocapside (génétique)</term><term>Protéines nucléocapside (métabolisme)</term><term>Protéines recombinantes ()</term><term>Protéines recombinantes 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Conformation</term><term>Protein Binding</term><term>Protein Structure, Tertiary</term><term>Regulatory Elements, Transcriptional</term><term>Sequence Homology, Amino Acid</term><term>Static Electricity</term><term>Thermodynamics</term></keywords><keywords scheme="MESH" xml:lang="fr"><term>ARN viral</term><term>Animaux</term><term>Cinétique</term><term>Conformation d'acide nucléique</term><term>Cristallographie aux rayons X</term><term>Données de séquences moléculaires</term><term>Liaison aux protéines</term><term>Modèles moléculaires</term><term>Mutagenèse dirigée</term><term>Protéines nucléocapside</term><term>Protéines recombinantes</term><term>Similitude de séquences d'acides aminés</term><term>Souris</term><term>Structure tertiaire des protéines</term><term>Séquence d'acides aminés</term><term>Séquence nucléotidique</term><term>Thermodynamique</term><term>Électricité statique</term><term>Éléments de régulation transcriptionnelle</term></keywords></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en">All coronaviruses (CoVs), including the causative agent of severe acute respiratory syndrome (SARS), encode a nucleocapsid (N) protein that harbors two independent RNA binding domains of known structure, but poorly characterized RNA binding properties. We show here that the N-terminal domain (NTD) of N protein from mouse hepatitis virus (MHV), a virus most closely related to SARS-CoV, employs aromatic amino acid-nucleobase stacking interactions with a triple adenosine motif to mediate high-affinity binding to single-stranded RNAs containing the transcriptional regulatory sequence (TRS) or its complement (cTRS). Stoichiometric NTD fully unwinds a TRS-cTRS duplex that mimics a transiently formed transcription intermediate in viral subgenomic RNA synthesis. Mutation of the solvent-exposed Y127, positioned on the beta-platform surface of our 1.75 A structure, binds the TRS far less tightly and is severely crippled in its RNA unwinding activity. In contrast, the C-terminal domain (CTD) exhibits no RNA unwinding activity. Viruses harboring Y127A N mutation are strongly selected against and Y127A N does not support an accessory function in MHV replication. We propose that the helix melting activity of the coronavirus N protein NTD plays a critical accessory role in subgenomic RNA synthesis and other processes requiring RNA remodeling.</div></front></TEI><affiliations><list><country><li>États-Unis</li></country><region><li>Indiana</li></region></list><tree><noCountry><name sortKey="Dann, Charles E" sort="Dann, Charles E" uniqKey="Dann C" first="Charles E" last="Dann">Charles E. Dann</name><name sortKey="Giedroc, David P" sort="Giedroc, David P" uniqKey="Giedroc D" first="David P" last="Giedroc">David P. Giedroc</name><name sortKey="Keane, Sarah C" sort="Keane, Sarah C" uniqKey="Keane S" first="Sarah C" last="Keane">Sarah C. Keane</name><name sortKey="Leibowitz, Julian L" sort="Leibowitz, Julian L" uniqKey="Leibowitz J" first="Julian L" last="Leibowitz">Julian L. Leibowitz</name><name sortKey="Li, Lichun" sort="Li, Lichun" uniqKey="Li L" first="Lichun" last="Li">Lichun Li</name><name sortKey="Liu, Pinghua" sort="Liu, Pinghua" uniqKey="Liu P" first="Pinghua" last="Liu">Pinghua Liu</name></noCountry><country name="États-Unis"><region name="Indiana"><name sortKey="Grossoehme, Nicholas E" sort="Grossoehme, Nicholas E" uniqKey="Grossoehme N" first="Nicholas E" last="Grossoehme">Nicholas E. Grossoehme</name></region></country></tree></affiliations></record>Pour manipuler ce document sous Unix (Dilib)
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