PMID- 19782089
OWN - NLM
STAT- MEDLINE
DCOM- 20091203
LR  - 20221207
IS  - 1089-8638 (Electronic)
IS  - 0022-2836 (Print)
IS  - 0022-2836 (Linking)
VI  - 394
IP  - 3
DP  - 2009 Dec 4
TI  - Coronavirus N protein N-terminal domain (NTD) specifically binds the 
      transcriptional regulatory sequence (TRS) and melts TRS-cTRS RNA duplexes.
PG  - 544-57
LID - 10.1016/j.jmb.2009.09.040 [doi]
AB  - 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.
FAU - Grossoehme, Nicholas E
AU  - Grossoehme NE
AD  - Department of Chemistry, Indiana University, Bloomington, IN 47405-7102, USA.
FAU - Li, Lichun
AU  - Li L
FAU - Keane, Sarah C
AU  - Keane SC
FAU - Liu, Pinghua
AU  - Liu P
FAU - Dann, Charles E 3rd
AU  - Dann CE 3rd
FAU - Leibowitz, Julian L
AU  - Leibowitz JL
FAU - Giedroc, David P
AU  - Giedroc DP
LA  - eng
SI  - PDB/3HD4
GR  - R01 AI067416/AI/NIAID NIH HHS/United States
GR  - R01 AI067416-01A1/AI/NIAID NIH HHS/United States
GR  - R01 AI067416-02/AI/NIAID NIH HHS/United States
GR  - R01 AI040187-11/AI/NIAID NIH HHS/United States
GR  - R01 AI040187-10/AI/NIAID NIH HHS/United States
GR  - R01 AI040187/AI/NIAID NIH HHS/United States
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
DEP - 20090924
PL  - Netherlands
TA  - J Mol Biol
JT  - Journal of molecular biology
JID - 2985088R
RN  - 0 (Coronavirus Nucleocapsid Proteins)
RN  - 0 (Nucleocapsid Proteins)
RN  - 0 (RNA, Viral)
RN  - 0 (Recombinant Proteins)
SB  - IM
MH  - Amino Acid Sequence
MH  - Animals
MH  - Base Sequence
MH  - Coronavirus Nucleocapsid Proteins
MH  - Crystallography, X-Ray
MH  - Kinetics
MH  - Mice
MH  - Models, Molecular
MH  - Molecular Sequence Data
MH  - Murine hepatitis virus/genetics/metabolism
MH  - Mutagenesis, Site-Directed
MH  - Nucleic Acid Conformation
MH  - Nucleocapsid Proteins/*chemistry/genetics/*metabolism
MH  - Protein Binding
MH  - Protein Structure, Tertiary
MH  - RNA, Viral/*chemistry/genetics/*metabolism
MH  - Recombinant Proteins/chemistry/genetics/metabolism
MH  - Regulatory Elements, Transcriptional
MH  - Severe acute respiratory syndrome-related coronavirus/genetics/metabolism
MH  - Sequence Homology, Amino Acid
MH  - Static Electricity
MH  - Thermodynamics
PMC - PMC2783395
MID - NIHMS149278
EDAT- 2009/09/29 06:00
MHDA- 2009/12/16 06:00
PMCR- 2009/09/24
CRDT- 2009/09/29 06:00
PHST- 2009/08/01 00:00 [received]
PHST- 2009/09/11 00:00 [revised]
PHST- 2009/09/16 00:00 [accepted]
PHST- 2009/09/29 06:00 [entrez]
PHST- 2009/09/29 06:00 [pubmed]
PHST- 2009/12/16 06:00 [medline]
PHST- 2009/09/24 00:00 [pmc-release]
AID - S0022-2836(09)01167-X [pii]
AID - 10.1016/j.jmb.2009.09.040 [doi]
PST - ppublish
SO  - J Mol Biol. 2009 Dec 4;394(3):544-57. doi: 10.1016/j.jmb.2009.09.040. Epub 2009 
      Sep 24.