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Journal of Virology, March 2008, p. 2161-2169, Vol. 82, No. 5
0022-538X/08/$08.00+0     doi:10.1128/JVI.01971-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.

The Vaccinia Virus B5 Protein Requires A34 for Efficient Intracellular Trafficking from the Endoplasmic Reticulum to the Site of Wrapping and Incorporation into Progeny Virions{triangledown}

Amalia K. Earley, Winnie M. Chan, and Brian M. Ward*

Department of Microbiology and Immunology, University of Rochester Medical Center, Rochester, New York 14642

Received 7 September 2007/ Accepted 4 December 2007

The glycoproteins encoded by the vaccinia virus A34R and B5R genes are involved in intracellular envelope virus formation and are highly conserved among orthopoxviruses. A recombinant virus that has the A34R gene deleted and the B5R gene replaced with a B5R gene fused to the enhanced green fluorescent protein (B5R-GFP) gene was created (vB5R-GFP/{Delta}A34R) to investigate the role of A34 during virion morphogenesis. Cells infected with vB5R-GFP/{Delta}A34R displayed GFP fluorescence throughout the cytoplasm, which differed markedly from that seen in cells infected with a normal B5R-GFP-expressing virus (vB5R-GFP). Immunofluorescence and subcellular fractionation demonstrated that B5-GFP localizes with the endoplasmic reticulum in the absence of A34. Expression of either full-length A34 or a construct consisting of the lumenal and transmembrane domains restored normal trafficking of B5-GFP to the site of wrapping in the juxtanuclear region. Coimmunoprecipitation studies confirmed that B5 and A34 interact through their luminal domains, and further analysis revealed that in the absence of A34, B5 is not efficiently incorporated into virions released from the cell.

* Corresponding author. Mailing address: Department of Microbiology and Immunology, University of Rochester Medical Center, 601 Elmwood Ave., Box 672, Rochester, NY 14642. Phone: (585) 275-9715. Fax: (585) 473-9573. E-mail: Brian_Ward{at}urmc.rochester.edu

{triangledown} Published ahead of print on 19 December 2007.

Journal of Virology, March 2008, p. 2161-2169, Vol. 82, No. 5
0022-538X/08/$08.00+0     doi:10.1128/JVI.01971-07
Copyright © 2008, American Society for Microbiology. All Rights Reserved.



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