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
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Steinfeldt, T.
Right arrow Articles by Mankertz, A.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Steinfeldt, T.
Right arrow Articles by Mankertz, A.

 Previous Article  |  Next Article 

Journal of Virology, June 2007, p. 5696-5704, Vol. 81, No. 11
0022-538X/07/$08.00+0     doi:10.1128/JVI.02420-06
Copyright © 2007, American Society for Microbiology. All Rights Reserved.

Functional Analysis of cis- and trans-Acting Replication Factors of Porcine Circovirus Type 1{triangledown}

Tobias Steinfeldt,2 Tim Finsterbusch,1 and Annette Mankertz1*

Division of Viral Infections (FG12), Robert Koch Institute, 13353 Berlin, Germany,1 Institute for Genetics, University of Cologne, 50674 Cologne, Germany2

Received 3 November 2006/ Accepted 6 March 2007

The replication proteins Rep and Rep' of porcine circovirus type 1 (PCV1) are both capable of introducing and resealing strand discontinuities at the viral origin of DNA replication in vitro underlying genome amplification by rolling-circle replication. The PCV1 origin of replication encompasses the minimal binding site (MBS) of the Rep and Rep' proteins and an inverted repeat with the potential to form a stem-loop. In this study, both elements of the PCV1 origin were demonstrated to be essential for viral replication in transfected cells. Furthermore, investigation of conserved amino acid motifs within Rep and Rep' proteins revealed that the mutation of motifs I, II, and III and of the GKS box interfered with viral replication. In vitro studies demonstrated that motifs I to III were essential for origin cleavage, while the GKS box was dispensable for the initiation of viral replication. A covalent link between Rep/Rep' and the DNA after origin cleavage was demonstrated, providing a mechanism for energy conservation for the termination of replication.

* Corresponding author. Mailing address: Division of Viral Infections (FG12), Robert Koch Institute, Nordufer 20, 13353 Berlin, Germany. Phone: 49 (0)30 4547 2516. Fax: 49 (0)30 4547 2598. E-mail: A.Mankertz{at}rki.de

{triangledown} Published ahead of print on 14 March 2007.

Journal of Virology, June 2007, p. 5696-5704, Vol. 81, No. 11
0022-538X/07/$08.00+0     doi:10.1128/JVI.02420-06
Copyright © 2007, 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»