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 HighWire
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow Articles by Lachmann, S.
Right arrow Articles by Nüesch, J. P. F.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Lachmann, S.
Right arrow Articles by Nüesch, J. P. F.

 Previous Article  |  Next Article 

Journal of Virology, July 2003, p. 8048-8060, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.8048-8060.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Novel PKC{eta} Is Required To Activate Replicative Functions of the Major Nonstructural Protein NS1 of Minute Virus of Mice

Sylvie Lachmann, Jean Rommeleare, and Jürg P. F. Nüesch*

Applied Tumour Virology Program, Department F010 and Institut National de la Santé et de la Recherche Médicale U375, Deutsches Krebsforschungszentrum, Heidelberg, Germany

Received 10 February 2003/ Accepted 1 May 2003

The multifunctional protein NS1 of minute virus of mice (MVMp) is posttranslationally modified and at least in part regulated by phosphorylation. The atypical lambda isoform of protein kinase C (PKC{lambda}) phosphorylates residues T435 and S473 in vitro and in vivo, leading directly to an activation of NS1 helicase function, but it is insufficient to activate NS1 for rolling circle replication. The present study identifies an additional cellular protein kinase phosphorylating and regulating NS1 activities. We show in vitro that the recombinant novel PKC{eta} phosphorylates NS1 and in consequence is able to activate the viral polypeptide in concert with PKC{lambda} for rolling circle replication. Moreover, this role of PKC{eta} was confirmed in vivo. We thereby created stably transfected A9 mouse fibroblasts, a typical MVMp-permissive host cell line with Flag-tagged constitutively active or inactive PKC{eta} mutants, in order to alter the activity of the NS1 regulating kinase. Indeed, tryptic phosphopeptide analyses of metabolically 32P-labeled NS1 expressed in the presence of a dominant-negative mutant, PKC{eta}DN, showed a lack of distinct NS1 phosphorylation events. This correlates with impaired synthesis of viral DNA replication intermediates, as detected by Southern blotting at the level of the whole cell population and by BrdU incorporation at the single-cell level. Remarkably, MVM infection triggers an accumulation of endogenous PKC{eta} in the nuclear periphery, suggesting that besides being a target for PKC{eta}, parvovirus infections may also affect the regulation of this NS1 regulating kinase. Altogether, our results underline the tight interconnection between PKC-mediated signaling and the parvoviral life cycle.

* Corresponding author. Mailing address: Applied Tumor Virology, Dept. F010 and Inserm U375, Deutsches Krebsforschungszentrum, Im Neuenheimer Feld 242, 69120 Heidelberg, Germany. Phone: 49-6221-424969. Fax: 49-6221-424962. E-mail: jpf.nuesch{at}dkfz.de.

Journal of Virology, July 2003, p. 8048-8060, Vol. 77, No. 14
0022-538X/03/$08.00+0     DOI: 10.1128/JVI.77.14.8048-8060.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Nuesch, J. P. F., Bar, S., Lachmann, S., Rommelaere, J. (2009). Ezrin-Radixin-Moesin Family Proteins Are Involved in Parvovirus Replication and Spreading. J. Virol. 83: 5854-5863 [Abstract] [Full Text]  
  • Nuesch, J. P. F., Rommelaere, J. (2007). A viral adaptor protein modulating casein kinase II activity induces cytopathic effects in permissive cells. Proc. Natl. Acad. Sci. USA 104: 12482-12487 [Abstract] [Full Text]  
  • Nuesch, J. P. F., Rommelaere, J. (2006). NS1 Interaction with CKII{alpha}: Novel Protein Complex Mediating Parvovirus-Induced Cytotoxicity.. J. Virol. 80: 4729-4739 [Abstract] [Full Text]  
  • Li, J., Werner, E., Hergenhahn, M., Poirey, R., Luo, Z., Rommelaere, J., Jauniaux, J.-C. (2005). Expression Profiling of Human Hepatoma Cells Reveals Global Repression of Genes Involved in Cell Proliferation, Growth, and Apoptosis upon Infection with Parvovirus H-1. J. Virol. 79: 2274-2286 [Abstract] [Full Text]  
  • Daeffler, L., Horlein, R., Rommelaere, J., Nuesch, J. P. F. (2003). Modulation of Minute Virus of Mice Cytotoxic Activities through Site-Directed Mutagenesis within the NS Coding Region. J. Virol. 77: 12466-12478 [Abstract] [Full Text]  


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