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Title: The nonstructural protein 8 (nsp8) of the SARS coronavirus interacts with its ORF6 accessory protein
Authors: Kumar, P.
Gunalan, V.
Liu, B. 
Chow, V.T.K. 
Druce, J.
Birch, C.
Catton, M.
Fielding, B.C.
Tan, Y.-J. 
Lal, S.K.
Keywords: Accessory protein
Non structural protein
Protein-protein interaction
SARS coronavirus
Yeast two-hybrid system
Issue Date: 30-Sep-2007
Source: Kumar, P., Gunalan, V., Liu, B., Chow, V.T.K., Druce, J., Birch, C., Catton, M., Fielding, B.C., Tan, Y.-J., Lal, S.K. (2007-09-30). The nonstructural protein 8 (nsp8) of the SARS coronavirus interacts with its ORF6 accessory protein. Virology 366 (2) : 293-303. ScholarBank@NUS Repository.
Abstract: 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. © 2007 Elsevier Inc. All rights reserved.
Source Title: Virology
ISSN: 00426822
DOI: 10.1016/j.virol.2007.04.029
Appears in Collections:Staff Publications

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