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Journal of Virology, July 2009, p. 6957-6962, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.00254-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

Detection of Nonstructural Protein 6 in Murine Coronavirus-Infected Cells and Analysis of the Transmembrane Topology by Using Bioinformatics and Molecular Approaches{triangledown}

Surendranath Baliji,1 Stephen A. Cammer,2 Bruno Sobral,2 and Susan C. Baker1*

Department of Microbiology and Immunology, Loyola University Stritch School of Medicine, Maywood, Illinois 60153,1 Virginia Bioinformatics Institute, Virginia Polytechnic Institute and State University, Blacksburg, Virginia 240612

Received 5 February 2009/ Accepted 13 April 2009

Coronaviruses encode large replicase polyproteins which are proteolytically processed by viral proteases to generate mature nonstructural proteins (nsps) that form the viral replication complex. Mouse hepatitis virus (MHV) replicase products nsp3, nsp4, and nsp6 are predicted to act as membrane anchors during assembly of the viral replication complexes. We report the first antibody-mediated Western blot detection of nsp6 from MHV-infected cells. The nsp6-specific peptide antiserum detected the replicase intermediate p150 (nsp4 to nsp11) and two nsp6 products of approximately 23 and 25 kDa. Analysis of nsp6 transmembrane topology revealed six membrane-spanning segments and a conserved hydrophobic domain in the C-terminal cytosolic tail.


* Corresponding author. Mailing address: Department of Microbiology & Immunology, Loyola University Medical Center, 2160 South First Ave., Bldg. 105, Maywood, IL 60153. Phone: (708) 216-6910. Fax: (708) 216-9574. E-mail: sbaker1{at}lumc.edu

{triangledown} Published ahead of print on 22 April 2009.


Journal of Virology, July 2009, p. 6957-6962, Vol. 83, No. 13
0022-538X/09/$08.00+0     doi:10.1128/JVI.00254-09
Copyright © 2009, American Society for Microbiology. All Rights Reserved.