Cargando…
Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes
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) o...
Autores principales: | , , , , , , |
---|---|
Formato: | Texto |
Lenguaje: | English |
Publicado: |
Elsevier Ltd.
2009
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2783395/ https://www.ncbi.nlm.nih.gov/pubmed/19782089 http://dx.doi.org/10.1016/j.jmb.2009.09.040 |
_version_ | 1782174694269517824 |
---|---|
author | Grossoehme, Nicholas E. Li, Lichun Keane, Sarah C. Liu, Pinghua Dann, Charles E. Leibowitz, Julian L. Giedroc, David P. |
author_facet | Grossoehme, Nicholas E. Li, Lichun Keane, Sarah C. Liu, Pinghua Dann, Charles E. Leibowitz, Julian L. Giedroc, David P. |
author_sort | Grossoehme, Nicholas E. |
collection | PubMed |
description | 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 β-platform surface of our 1.75 Å 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. |
format | Text |
id | pubmed-2783395 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | Elsevier Ltd. |
record_format | MEDLINE/PubMed |
spelling | pubmed-27833952010-12-07 Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes Grossoehme, Nicholas E. Li, Lichun Keane, Sarah C. Liu, Pinghua Dann, Charles E. Leibowitz, Julian L. Giedroc, David P. J Mol Biol Article 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 β-platform surface of our 1.75 Å 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. Elsevier Ltd. 2009-12-04 2009-09-24 /pmc/articles/PMC2783395/ /pubmed/19782089 http://dx.doi.org/10.1016/j.jmb.2009.09.040 Text en Copyright © 2009 Elsevier Ltd. All rights reserved. Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active. |
spellingShingle | Article Grossoehme, Nicholas E. Li, Lichun Keane, Sarah C. Liu, Pinghua Dann, Charles E. Leibowitz, Julian L. Giedroc, David P. Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes |
title | Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes |
title_full | Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes |
title_fullStr | Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes |
title_full_unstemmed | Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes |
title_short | Coronavirus N Protein N-Terminal Domain (NTD) Specifically Binds the Transcriptional Regulatory Sequence (TRS) and Melts TRS-cTRS RNA Duplexes |
title_sort | coronavirus n protein n-terminal domain (ntd) specifically binds the transcriptional regulatory sequence (trs) and melts trs-ctrs rna duplexes |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2783395/ https://www.ncbi.nlm.nih.gov/pubmed/19782089 http://dx.doi.org/10.1016/j.jmb.2009.09.040 |
work_keys_str_mv | AT grossoehmenicholase coronavirusnproteinnterminaldomainntdspecificallybindsthetranscriptionalregulatorysequencetrsandmeltstrsctrsrnaduplexes AT lilichun coronavirusnproteinnterminaldomainntdspecificallybindsthetranscriptionalregulatorysequencetrsandmeltstrsctrsrnaduplexes AT keanesarahc coronavirusnproteinnterminaldomainntdspecificallybindsthetranscriptionalregulatorysequencetrsandmeltstrsctrsrnaduplexes AT liupinghua coronavirusnproteinnterminaldomainntdspecificallybindsthetranscriptionalregulatorysequencetrsandmeltstrsctrsrnaduplexes AT danncharlese coronavirusnproteinnterminaldomainntdspecificallybindsthetranscriptionalregulatorysequencetrsandmeltstrsctrsrnaduplexes AT leibowitzjulianl coronavirusnproteinnterminaldomainntdspecificallybindsthetranscriptionalregulatorysequencetrsandmeltstrsctrsrnaduplexes AT giedrocdavidp coronavirusnproteinnterminaldomainntdspecificallybindsthetranscriptionalregulatorysequencetrsandmeltstrsctrsrnaduplexes |