Multiple nucleic acid binding sites and intrinsic disorder of severe acute respiratory syndrome coronavirus nucleocapsid protein: Implications for ribonucleocapsid protein packaging

Chung Ke Chang, Yen Lan Hsu, Yuan Hsiang Chang, Fa An Chao, Ming Chya Wu, Yu Shan Huang, Chin Kun Hu, Tai Huang Huang

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

The nucleocapsid protein (N) of the severe acute respiratory syndrome coronavirus (SARS-CoV) packages the viral genomic RNA and is crucial for viability. However, the RNA-binding mechanism is poorly understood. We have shown previously that the N protein contains two structural domains-the N-terminal domain (NTD; residues 45 to 181) and the C-terminal dimerization domain (CTD; residues 248 to 365)-flanked by long stretches of disordered regions accounting for almost half of the entire sequence. Small-angle X-ray scattering data show that the protein is in an extended conformation and that the two structural domains of the SARS-CoV N protein are far apart. Both the NTD and the CTD have been shown to bind RNA. Here we show that all disordered regions are also capable of binding to RNA. Constructs containing multiple RNA-binding regions showed Hill coefficients greater than 1, suggesting that the N protein binds to RNA cooperatively. The effect can be explained by the "coupled-allostery" model, devised to explain the allosteric effect in a multido- main regulatory system. Although the N proteins of different coronaviruses share very low sequence homology, the physicochemical features described above may be conserved across different groups of Coronaviridae. The current results underscore the important roles of multisite nucleic acid binding and intrinsic disorder in N protein function and RNP packaging.

Original languageEnglish
Pages (from-to)2255-2264
Number of pages10
JournalJournal of Virology
Volume83
Issue number5
DOIs
Publication statusPublished - 2009 Mar 1

Fingerprint

Severe Acute Respiratory Syndrome
Product Packaging
Nucleic Acids
Binding Sites
RNA
Proteins
Coronaviridae
Nucleocapsid Proteins
Coronavirus
Viral RNA
Dimerization
Sequence Homology
X-Rays
Coronavirus nucleocapsid protein

ASJC Scopus subject areas

  • Microbiology
  • Immunology
  • Insect Science
  • Virology

Cite this

Multiple nucleic acid binding sites and intrinsic disorder of severe acute respiratory syndrome coronavirus nucleocapsid protein : Implications for ribonucleocapsid protein packaging. / Chang, Chung Ke; Hsu, Yen Lan; Chang, Yuan Hsiang; Chao, Fa An; Wu, Ming Chya; Huang, Yu Shan; Hu, Chin Kun; Huang, Tai Huang.

In: Journal of Virology, Vol. 83, No. 5, 01.03.2009, p. 2255-2264.

Research output: Contribution to journalArticle

Chang, Chung Ke ; Hsu, Yen Lan ; Chang, Yuan Hsiang ; Chao, Fa An ; Wu, Ming Chya ; Huang, Yu Shan ; Hu, Chin Kun ; Huang, Tai Huang. / Multiple nucleic acid binding sites and intrinsic disorder of severe acute respiratory syndrome coronavirus nucleocapsid protein : Implications for ribonucleocapsid protein packaging. In: Journal of Virology. 2009 ; Vol. 83, No. 5. pp. 2255-2264.
@article{905892f5479b412a9db617b85a002277,
title = "Multiple nucleic acid binding sites and intrinsic disorder of severe acute respiratory syndrome coronavirus nucleocapsid protein: Implications for ribonucleocapsid protein packaging",
abstract = "The nucleocapsid protein (N) of the severe acute respiratory syndrome coronavirus (SARS-CoV) packages the viral genomic RNA and is crucial for viability. However, the RNA-binding mechanism is poorly understood. We have shown previously that the N protein contains two structural domains-the N-terminal domain (NTD; residues 45 to 181) and the C-terminal dimerization domain (CTD; residues 248 to 365)-flanked by long stretches of disordered regions accounting for almost half of the entire sequence. Small-angle X-ray scattering data show that the protein is in an extended conformation and that the two structural domains of the SARS-CoV N protein are far apart. Both the NTD and the CTD have been shown to bind RNA. Here we show that all disordered regions are also capable of binding to RNA. Constructs containing multiple RNA-binding regions showed Hill coefficients greater than 1, suggesting that the N protein binds to RNA cooperatively. The effect can be explained by the {"}coupled-allostery{"} model, devised to explain the allosteric effect in a multido- main regulatory system. Although the N proteins of different coronaviruses share very low sequence homology, the physicochemical features described above may be conserved across different groups of Coronaviridae. The current results underscore the important roles of multisite nucleic acid binding and intrinsic disorder in N protein function and RNP packaging.",
author = "Chang, {Chung Ke} and Hsu, {Yen Lan} and Chang, {Yuan Hsiang} and Chao, {Fa An} and Wu, {Ming Chya} and Huang, {Yu Shan} and Hu, {Chin Kun} and Huang, {Tai Huang}",
year = "2009",
month = "3",
day = "1",
doi = "10.1128/JVI.02001-08",
language = "English",
volume = "83",
pages = "2255--2264",
journal = "Journal of Virology",
issn = "0022-538X",
publisher = "American Society for Microbiology",
number = "5",

}

TY - JOUR

T1 - Multiple nucleic acid binding sites and intrinsic disorder of severe acute respiratory syndrome coronavirus nucleocapsid protein

T2 - Implications for ribonucleocapsid protein packaging

AU - Chang, Chung Ke

AU - Hsu, Yen Lan

AU - Chang, Yuan Hsiang

AU - Chao, Fa An

AU - Wu, Ming Chya

AU - Huang, Yu Shan

AU - Hu, Chin Kun

AU - Huang, Tai Huang

PY - 2009/3/1

Y1 - 2009/3/1

N2 - The nucleocapsid protein (N) of the severe acute respiratory syndrome coronavirus (SARS-CoV) packages the viral genomic RNA and is crucial for viability. However, the RNA-binding mechanism is poorly understood. We have shown previously that the N protein contains two structural domains-the N-terminal domain (NTD; residues 45 to 181) and the C-terminal dimerization domain (CTD; residues 248 to 365)-flanked by long stretches of disordered regions accounting for almost half of the entire sequence. Small-angle X-ray scattering data show that the protein is in an extended conformation and that the two structural domains of the SARS-CoV N protein are far apart. Both the NTD and the CTD have been shown to bind RNA. Here we show that all disordered regions are also capable of binding to RNA. Constructs containing multiple RNA-binding regions showed Hill coefficients greater than 1, suggesting that the N protein binds to RNA cooperatively. The effect can be explained by the "coupled-allostery" model, devised to explain the allosteric effect in a multido- main regulatory system. Although the N proteins of different coronaviruses share very low sequence homology, the physicochemical features described above may be conserved across different groups of Coronaviridae. The current results underscore the important roles of multisite nucleic acid binding and intrinsic disorder in N protein function and RNP packaging.

AB - The nucleocapsid protein (N) of the severe acute respiratory syndrome coronavirus (SARS-CoV) packages the viral genomic RNA and is crucial for viability. However, the RNA-binding mechanism is poorly understood. We have shown previously that the N protein contains two structural domains-the N-terminal domain (NTD; residues 45 to 181) and the C-terminal dimerization domain (CTD; residues 248 to 365)-flanked by long stretches of disordered regions accounting for almost half of the entire sequence. Small-angle X-ray scattering data show that the protein is in an extended conformation and that the two structural domains of the SARS-CoV N protein are far apart. Both the NTD and the CTD have been shown to bind RNA. Here we show that all disordered regions are also capable of binding to RNA. Constructs containing multiple RNA-binding regions showed Hill coefficients greater than 1, suggesting that the N protein binds to RNA cooperatively. The effect can be explained by the "coupled-allostery" model, devised to explain the allosteric effect in a multido- main regulatory system. Although the N proteins of different coronaviruses share very low sequence homology, the physicochemical features described above may be conserved across different groups of Coronaviridae. The current results underscore the important roles of multisite nucleic acid binding and intrinsic disorder in N protein function and RNP packaging.

UR - http://www.scopus.com/inward/record.url?scp=60049100937&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=60049100937&partnerID=8YFLogxK

U2 - 10.1128/JVI.02001-08

DO - 10.1128/JVI.02001-08

M3 - Article

C2 - 19052082

AN - SCOPUS:60049100937

VL - 83

SP - 2255

EP - 2264

JO - Journal of Virology

JF - Journal of Virology

SN - 0022-538X

IS - 5

ER -