This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Download to citation manager
Right arrowReprints and Permissions
Right arrow Copyright Information
Right arrow Books from ASM Press
Right arrow MicrobeWorld
Google Scholar
Right arrow Articles by Sperling, K. M.
Right arrow Articles by Sutter, G.
PubMed
Right arrow PubMed Citation
Right arrow Articles by Sperling, K. M.
Right arrow Articles by Sutter, G.

 Previous Article  |  Next Article 

Journal of Virology, June 2009, p. 6029-6038, Vol. 83, No. 12
0022-538X/09/$08.00+0     doi:10.1128/JVI.01628-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.

The Orthopoxvirus 68-Kilodalton Ankyrin-Like Protein Is Essential for DNA Replication and Complete Gene Expression of Modified Vaccinia Virus Ankara in Nonpermissive Human and Murine Cells{triangledown}

Karin M. Sperling,1 Astrid Schwantes,1 Caroline Staib,2 Barbara S. Schnierle,1 and Gerd Sutter1*

Paul-Ehrlich-Institut, Division of Virology, Paul-Ehrlich-Strasse 51-59, 63225 Langen, Germany,1 Institute of Virology, Technische Universität München, 81675 Munich, Germany2

Received 30 July 2008/ Accepted 29 March 2009

Modified vaccinia virus Ankara (MVA) is a highly attenuated and replication-deficient vaccinia virus (VACV) that is being evaluated as replacement smallpox vaccine and candidate viral vector. MVA lacks many genes associated with virulence and/or regulation of virus tropism. The 68-kDa ankyrin-like protein (68k-ank) is the only ankyrin repeat-containing protein that is encoded by the MVA genome and is highly conserved throughout the Orthopoxvirus genus. We showed previously that 68k-ank is composed of ankyrin repeats and an F-box-like domain and forms an SCF ubiquitin ligase complex together with the cellular proteins Skp1a and Cullin-1. We now report that 68k-ank (MVA open reading frame 186R) is an essential factor for completion of the MVA intracellular life cycle in nonpermissive human and murine cells. Infection of mouse NIH 3T3 and human HaCaT cells with MVA with a deletion of the 68k-ank gene (MVA-{Delta}68k-ank) was characterized by an extensive reduction of viral intermediate RNA and protein, as well as late transcripts and drastically impaired late protein synthesis. Furthermore, infections with MVA-{Delta}68k-ank failed to induce the host protein shutoff that is characteristic of VACV infections. Although we demonstrated that proteasome function in general is essential for the completion of the MVA molecular life cycle, we found that a mutant 68k-ank protein with a deletion of the F-box-like domain was able to fully complement the deficiency of MVA-{Delta}68k-ank to express all classes of viral genes. Thus, our data demonstrate that the 68k-ank protein contains another critical domain that may function independently of SCF ubiquitin ligase complex formation, suggesting multiple activities of this interesting regulatory protein.

* Corresponding author. Mailing address: Paul-Ehrlich-Institut, Paul-Ehrlich-Strasse 51-59, 63225 Langen, Germany. Phone: 49 6103 77 2140. Fax: 49 6103 77 1273. E-mail: sutge{at}pei.de

{triangledown} Published ahead of print on 8 April 2009.

Journal of Virology, June 2009, p. 6029-6038, Vol. 83, No. 12
0022-538X/09/$08.00+0     doi:10.1128/JVI.01628-08
Copyright © 2009, American Society for Microbiology. All Rights Reserved.





Copyright © 2009 by the American Society for Microbiology.   For an alternate route to Journals.ASM.org, visit: http://intl-journals.asm.org | More Info»