Source:http://linkedlifedata.com/resource/pubmed/id/17532020
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rdf:type | |
lifeskim:mentions | |
pubmed:issue |
2
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pubmed:dateCreated |
2007-9-18
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pubmed:abstractText |
Severe acute respiratory syndrome (SARS) coronavirus (SARS-CoV) caused a severe outbreak in several regions of the world in 2003. The SARS-CoV genome is predicted to contain 14 functional open reading frames (ORFs). The first ORF (1a and 1b) encodes a large polyprotein that is cleaved into nonstructural proteins (nsp). The other ORFs encode for four structural proteins (spike, membrane, nucleocapsid and envelope) as well as eight SARS-CoV-specific accessory proteins (3a, 3b, 6, 7a, 7b, 8a, 8b and 9b). In this report we have cloned the predicted nsp8 gene and the ORF6 gene of the SARS-CoV and studied their abilities to interact with each other. We expressed the two proteins as fusion proteins in the yeast two-hybrid system to demonstrate protein-protein interactions and tested the same using a yeast genetic cross. Further the strength of the interaction was measured by challenging growth of the positive interaction clones on increasing gradients of 2-amino trizole. The interaction was then verified by expressing both proteins separately in-vitro in a coupled-transcription translation system and by coimmunoprecipitation in mammalian cells. Finally, colocalization experiments were performed in SARS-CoV infected Vero E6 mammalian cells to confirm the nsp8-ORF6 interaction. To the best of our knowledge, this is the first report of the interaction between a SARS-CoV accessory protein and nsp8 and our findings suggest that ORF6 protein may play a role in virus replication.
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pubmed:language |
eng
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pubmed:journal | |
pubmed:citationSubset |
IM
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pubmed:chemical | |
pubmed:status |
MEDLINE
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pubmed:month |
Sep
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pubmed:issn |
0042-6822
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pubmed:author | |
pubmed:issnType |
Print
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pubmed:day |
30
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pubmed:volume |
366
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pubmed:owner |
NLM
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pubmed:authorsComplete |
Y
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pubmed:pagination |
293-303
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pubmed:meshHeading |
pubmed-meshheading:17532020-Animals,
pubmed-meshheading:17532020-Cell Line,
pubmed-meshheading:17532020-Cercopithecus aethiops,
pubmed-meshheading:17532020-Cloning, Molecular,
pubmed-meshheading:17532020-Cytoplasm,
pubmed-meshheading:17532020-Immunoprecipitation,
pubmed-meshheading:17532020-Microscopy, Confocal,
pubmed-meshheading:17532020-Microscopy, Fluorescence,
pubmed-meshheading:17532020-Protein Binding,
pubmed-meshheading:17532020-SARS Virus,
pubmed-meshheading:17532020-Two-Hybrid System Techniques,
pubmed-meshheading:17532020-Viral Nonstructural Proteins
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pubmed:year |
2007
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pubmed:articleTitle |
The nonstructural protein 8 (nsp8) of the SARS coronavirus interacts with its ORF6 accessory protein.
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pubmed:affiliation |
Virology Group, International Centre for Genetic Engineering and Biotechnology, P. O. Box: 10504, Aruna Asaf Ali Road, New Delhi 110067, India.
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pubmed:publicationType |
Journal Article,
Research Support, Non-U.S. Gov't
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