The nucleocapsid protein of SARS coronavirus has a high binding affinity to the human cellular heterogeneous nuclear ribonucleoprotein A1.
Identifieur interne : 004589 ( Main/Exploration ); précédent : 004588; suivant : 004590The nucleocapsid protein of SARS coronavirus has a high binding affinity to the human cellular heterogeneous nuclear ribonucleoprotein A1.
Auteurs : Haibin Luo [République populaire de Chine] ; Qing Chen ; Jing Chen ; Kaixian Chen ; Xu Shen ; Hualiang JiangSource :
- FEBS letters [ 0014-5793 ] ; 2005.
Descripteurs français
- KwdFr :
- ARN viral (biosynthèse), Cinétique, Clones cellulaires, Fragments peptidiques (métabolisme), Glutathione transferase (métabolisme), Glycine (), Humains, Protéines nucléocapside (), Protéines nucléocapside (génétique), Protéines nucléocapside (isolement et purification), Protéines nucléocapside (métabolisme), Protéines recombinantes (métabolisme), Ribonucléoprotéine nucléaire hétérogène du groupe A-B (), Ribonucléoprotéine nucléaire hétérogène du groupe A-B (métabolisme), Résonance plasmonique de surface, Saccharomyces cerevisiae (croissance et développement), Saccharomyces cerevisiae (métabolisme), Structure tertiaire des protéines, Syndrome respiratoire aigu sévère (métabolisme), Techniques de double hybride, Transformation génétique, Électrophorèse sur gel de polyacrylamide.
- MESH :
- biosynthèse : ARN viral.
- croissance et développement : Saccharomyces cerevisiae.
- génétique : Protéines nucléocapside.
- isolement et purification : Protéines nucléocapside.
- métabolisme : Fragments peptidiques, Glutathione transferase, Protéines nucléocapside, Protéines recombinantes, Ribonucléoprotéine nucléaire hétérogène du groupe A-B, Saccharomyces cerevisiae, Syndrome respiratoire aigu sévère.
- Cinétique, Clones cellulaires, Glycine, Humains, Protéines nucléocapside, Ribonucléoprotéine nucléaire hétérogène du groupe A-B, Résonance plasmonique de surface, Structure tertiaire des protéines, Techniques de double hybride, Transformation génétique, Électrophorèse sur gel de polyacrylamide.
English descriptors
- KwdEn :
- Clone Cells, Electrophoresis, Polyacrylamide Gel, Glutathione Transferase (metabolism), Glycine (chemistry), Heterogeneous Nuclear Ribonucleoprotein A1, Heterogeneous-Nuclear Ribonucleoprotein Group A-B (chemistry), Heterogeneous-Nuclear Ribonucleoprotein Group A-B (metabolism), Humans, Kinetics, Nucleocapsid Proteins (chemistry), Nucleocapsid Proteins (genetics), Nucleocapsid Proteins (isolation & purification), Nucleocapsid Proteins (metabolism), Peptide Fragments (metabolism), Protein Structure, Tertiary, RNA, Viral (biosynthesis), Recombinant Proteins (metabolism), Saccharomyces cerevisiae (growth & development), Saccharomyces cerevisiae (metabolism), Severe Acute Respiratory Syndrome (metabolism), Surface Plasmon Resonance, Transformation, Genetic, Two-Hybrid System Techniques.
- MESH :
- chemical , biosynthesis : RNA, Viral.
- chemical , chemistry : Glycine, Heterogeneous-Nuclear Ribonucleoprotein Group A-B, Nucleocapsid Proteins.
- chemical , genetics : Nucleocapsid Proteins.
- chemical , isolation & purification : Nucleocapsid Proteins.
- chemical , metabolism : Glutathione Transferase, Heterogeneous-Nuclear Ribonucleoprotein Group A-B, Nucleocapsid Proteins, Peptide Fragments, Recombinant Proteins.
- growth & development : Saccharomyces cerevisiae.
- metabolism : Saccharomyces cerevisiae, Severe Acute Respiratory Syndrome.
- Clone Cells, Electrophoresis, Polyacrylamide Gel, Heterogeneous Nuclear Ribonucleoprotein A1, Humans, Kinetics, Protein Structure, Tertiary, Surface Plasmon Resonance, Transformation, Genetic, Two-Hybrid System Techniques.
Abstract
The nucleocapsid (N) protein of SARS coronavirus (SARS_CoV) is a major structural component of virions, which appears to be a multifunctional protein involved in viral RNA replication and translation. Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is related to the pre-mRNA splicing in the nucleus and translation regulation in the cytoplasm. In this report, based on the relevant biophysical and biochemical assays, the nucleocapsid protein of SARS_CoV (SARS_N) was discovered to exhibit high binding affinity to human hnRNP A1. GST pull-down results clearly demonstrated that SARS_N protein could directly and specifically bind to human hnRNP A1 in vitro. Yeast two-hybrid assays further indicated in vivo that such binding relates to the fragment (aa 161-210) of SARS_N and the Gly-rich domain (aa 203-320) of hnRNP A1. Moreover, kinetic analyses by surface plasmon resonance (SPR) technology revealed that SARS_N protein has a specific binding affinity against human hnRNP A1 with K(D) at 0.35 +/- 0.02 microM (k(on) = 5.83 +/- 0.42 x 10(3) M(-1)s(-1) and k(off) = 2.06 +/- 0.12 x 10(-3)s(-1)). It is suggested that both SARS_N and hnRNP A1 proteins are possibly within the SARS_CoV replication/transcription complex and SARS_N/human hnRNP A1 interaction might function in the regulation of SARS_CoV RNA synthesis. In addition, the determined results showed that SARS_N protein has only one binding domain for interacting with human hnRNP A1, which is different from the mouse hepatitis virus (MHV) binding case where the nucleocapsid protein of MHV (MHV_N) was found to have two binding domains involved in the MHV_N/hnRNP A1 interaction, thereby suggesting that SARS_N protein might carry out a different binding mode to bind to human hnRNP A1 for its further function performance in comparison with MHV_N.
DOI: 10.1016/j.febslet.2005.03.080
PubMed: 15862300
Affiliations:
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Le document en format XML
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<term>Glycine (chemistry)</term>
<term>Heterogeneous Nuclear Ribonucleoprotein A1</term>
<term>Heterogeneous-Nuclear Ribonucleoprotein Group A-B (chemistry)</term>
<term>Heterogeneous-Nuclear Ribonucleoprotein Group A-B (metabolism)</term>
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<term>Saccharomyces cerevisiae (growth & development)</term>
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<term>Saccharomyces cerevisiae (métabolisme)</term>
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<term>Techniques de double hybride</term>
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<front><div type="abstract" xml:lang="en">The nucleocapsid (N) protein of SARS coronavirus (SARS_CoV) is a major structural component of virions, which appears to be a multifunctional protein involved in viral RNA replication and translation. Heterogeneous nuclear ribonucleoprotein A1 (hnRNP A1) is related to the pre-mRNA splicing in the nucleus and translation regulation in the cytoplasm. In this report, based on the relevant biophysical and biochemical assays, the nucleocapsid protein of SARS_CoV (SARS_N) was discovered to exhibit high binding affinity to human hnRNP A1. GST pull-down results clearly demonstrated that SARS_N protein could directly and specifically bind to human hnRNP A1 in vitro. Yeast two-hybrid assays further indicated in vivo that such binding relates to the fragment (aa 161-210) of SARS_N and the Gly-rich domain (aa 203-320) of hnRNP A1. Moreover, kinetic analyses by surface plasmon resonance (SPR) technology revealed that SARS_N protein has a specific binding affinity against human hnRNP A1 with K(D) at 0.35 +/- 0.02 microM (k(on) = 5.83 +/- 0.42 x 10(3) M(-1)s(-1) and k(off) = 2.06 +/- 0.12 x 10(-3)s(-1)). It is suggested that both SARS_N and hnRNP A1 proteins are possibly within the SARS_CoV replication/transcription complex and SARS_N/human hnRNP A1 interaction might function in the regulation of SARS_CoV RNA synthesis. In addition, the determined results showed that SARS_N protein has only one binding domain for interacting with human hnRNP A1, which is different from the mouse hepatitis virus (MHV) binding case where the nucleocapsid protein of MHV (MHV_N) was found to have two binding domains involved in the MHV_N/hnRNP A1 interaction, thereby suggesting that SARS_N protein might carry out a different binding mode to bind to human hnRNP A1 for its further function performance in comparison with MHV_N.</div>
</front>
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<name sortKey="Chen, Kaixian" sort="Chen, Kaixian" uniqKey="Chen K" first="Kaixian" last="Chen">Kaixian Chen</name>
<name sortKey="Chen, Qing" sort="Chen, Qing" uniqKey="Chen Q" first="Qing" last="Chen">Qing Chen</name>
<name sortKey="Jiang, Hualiang" sort="Jiang, Hualiang" uniqKey="Jiang H" first="Hualiang" last="Jiang">Hualiang Jiang</name>
<name sortKey="Shen, Xu" sort="Shen, Xu" uniqKey="Shen X" first="Xu" last="Shen">Xu Shen</name>
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<country name="République populaire de Chine"><noRegion><name sortKey="Luo, Haibin" sort="Luo, Haibin" uniqKey="Luo H" first="Haibin" last="Luo">Haibin Luo</name>
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