The cellular RNA helicase DDX1 interacts with coronavirus nonstructural protein 14 and enhances viral replication.
Identifieur interne : 002424 ( Main/Exploration ); précédent : 002423; suivant : 002425The cellular RNA helicase DDX1 interacts with coronavirus nonstructural protein 14 and enhances viral replication.
Auteurs : Linghui Xu [Singapour] ; Siti Khadijah ; Shouguo Fang ; Li Wang ; Felicia P L. Tay ; Ding Xiang LiuSource :
- Journal of virology [ 1098-5514 ] ; 2010.
Descripteurs français
- KwdFr :
- Animaux, Cellules HeLa, Cellules Vero, Cytoplasme (métabolisme), DEAD-box RNA helicases (génétique), DEAD-box RNA helicases (métabolisme), Humains, Infections à coronavirus (génétique), Infections à coronavirus (métabolisme), Infections à coronavirus (virologie), Liaison aux protéines, Noyau de la cellule (métabolisme), Protéines virales non structurales (génétique), Protéines virales non structurales (métabolisme), Réplication virale, Techniques de double hybride, Transport de protéines, Virus de la bronchite infectieuse (génétique), Virus de la bronchite infectieuse (physiologie).
- MESH :
- génétique : DEAD-box RNA helicases, Infections à coronavirus, Protéines virales non structurales, Virus de la bronchite infectieuse.
- métabolisme : Cytoplasme, DEAD-box RNA helicases, Infections à coronavirus, Noyau de la cellule, Protéines virales non structurales.
- physiologie : Virus de la bronchite infectieuse.
- virologie : Infections à coronavirus.
- Animaux, Cellules HeLa, Cellules Vero, Humains, Liaison aux protéines, Réplication virale, Techniques de double hybride, Transport de protéines.
English descriptors
- KwdEn :
- Animals, Cell Nucleus (metabolism), Chlorocebus aethiops, Coronavirus Infections (genetics), Coronavirus Infections (metabolism), Coronavirus Infections (virology), Cytoplasm (metabolism), DEAD-box RNA Helicases (genetics), DEAD-box RNA Helicases (metabolism), HeLa Cells, Humans, Infectious bronchitis virus (genetics), Infectious bronchitis virus (physiology), Protein Binding, Protein Transport, Two-Hybrid System Techniques, Vero Cells, Viral Nonstructural Proteins (genetics), Viral Nonstructural Proteins (metabolism), Virus Replication.
- MESH :
- chemical , genetics : DEAD-box RNA Helicases, Viral Nonstructural Proteins.
- genetics : Coronavirus Infections, Infectious bronchitis virus.
- metabolism : Cell Nucleus, Coronavirus Infections, Cytoplasm, DEAD-box RNA Helicases, Viral Nonstructural Proteins.
- physiology : Infectious bronchitis virus.
- virology : Coronavirus Infections.
- Animals, Chlorocebus aethiops, HeLa Cells, Humans, Protein Binding, Protein Transport, Two-Hybrid System Techniques, Vero Cells, Virus Replication.
Abstract
The involvement of host proteins in the replication and transcription of viral RNA is a poorly understood area for many RNA viruses. For coronaviruses, it was long speculated that replication of the giant RNA genome and transcription of multiple subgenomic mRNA species by a unique discontinuous transcription mechanism may require host cofactors. To search for such cellular proteins, yeast two-hybrid screening was carried out by using the nonstructural protein 14 (nsp14) from the coronavirus infectious bronchitis virus (IBV) as a bait protein, leading to the identification of DDX1, a cellular RNA helicase in the DExD/H helicase family, as a potential interacting partner. This interaction was subsequently confirmed by coimmunoprecipitation assays with cells coexpressing the two proteins and with IBV-infected cells. Furthermore, the endogenous DDX1 protein was found to be relocated from the nucleus to the cytoplasm in IBV-infected cells. In addition to its interaction with IBV nsp14, DDX1 could also interact with the nsp14 protein from severe acute respiratory syndrome coronavirus (SARS-CoV), suggesting that interaction with DDX1 may be a general feature of coronavirus nsp14. The interacting domains were mapped to the C-terminal region of DDX1 containing motifs V and VI and to the N-terminal portion of nsp14. Manipulation of DDX1 expression, either by small interfering RNA-induced knockdown or by overexpression of a mutant DDX1 protein, confirmed that this interaction may enhance IBV replication. This study reveals that DDX1 contributes to efficient coronavirus replication in cell culture.
DOI: 10.1128/JVI.00392-10
PubMed: 20573827
Affiliations:
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Le document en format XML
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<term>Coronavirus Infections (metabolism)</term>
<term>Coronavirus Infections (virology)</term>
<term>Cytoplasm (metabolism)</term>
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<term>DEAD-box RNA Helicases (metabolism)</term>
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<front><div type="abstract" xml:lang="en">The involvement of host proteins in the replication and transcription of viral RNA is a poorly understood area for many RNA viruses. For coronaviruses, it was long speculated that replication of the giant RNA genome and transcription of multiple subgenomic mRNA species by a unique discontinuous transcription mechanism may require host cofactors. To search for such cellular proteins, yeast two-hybrid screening was carried out by using the nonstructural protein 14 (nsp14) from the coronavirus infectious bronchitis virus (IBV) as a bait protein, leading to the identification of DDX1, a cellular RNA helicase in the DExD/H helicase family, as a potential interacting partner. This interaction was subsequently confirmed by coimmunoprecipitation assays with cells coexpressing the two proteins and with IBV-infected cells. Furthermore, the endogenous DDX1 protein was found to be relocated from the nucleus to the cytoplasm in IBV-infected cells. In addition to its interaction with IBV nsp14, DDX1 could also interact with the nsp14 protein from severe acute respiratory syndrome coronavirus (SARS-CoV), suggesting that interaction with DDX1 may be a general feature of coronavirus nsp14. The interacting domains were mapped to the C-terminal region of DDX1 containing motifs V and VI and to the N-terminal portion of nsp14. Manipulation of DDX1 expression, either by small interfering RNA-induced knockdown or by overexpression of a mutant DDX1 protein, confirmed that this interaction may enhance IBV replication. This study reveals that DDX1 contributes to efficient coronavirus replication in cell culture.</div>
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<name sortKey="Liu, Ding Xiang" sort="Liu, Ding Xiang" uniqKey="Liu D" first="Ding Xiang" last="Liu">Ding Xiang Liu</name>
<name sortKey="Tay, Felicia P L" sort="Tay, Felicia P L" uniqKey="Tay F" first="Felicia P L" last="Tay">Felicia P L. Tay</name>
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