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Journal of Virology, July 2005, p. 8422-8430, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8422-8430.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

A Functional Histidine-Tagged Replication Initiator Protein: Implications for the Study of Single-Stranded DNA Virus Replication In Planta{dagger}

Julio C. Vega-Arreguín, Tatiana Timchenko, Bruno Gronenborn,* and Bertha Cecilia Ramírez*

Institut des Sciences du Végétal, CNRS, 91198 Gif sur Yvette Cedex, France

Received 19 November 2004/ Accepted 6 March 2005

Replication initiation of nanoviruses, plant viruses with a multipartite circular single-stranded DNA genome, is triggered by the master Rep (M-Rep) protein. To enable the study of interactions between M-Rep and viral or host factors involved in replication, we designed oligohistidine-tagged variants of the nanovirus Faba bean necrotic yellows virus (FBNYV) M-Rep protein that allow affinity purification of enzymatically active M-Rep from plant tissue. The tagged M-Rep protein was able to initiate replication of its cognate and other FBNYV DNAs in Nicotiana benthamiana leaf disks and plants. The replicon encoding the tagged M-Rep protein multiplied and moved systemically in FBNYV-infected Vicia faba plants and was transmitted by the aphid vector of the virus. Using the tagged M-Rep protein, we demonstrated the in planta interaction between wild-type M-Rep and its tagged counterpart. Such a tagged and fully functional replication initiator protein will have bearings on the isolation of protein complexes from plants.

* Corresponding author. Mailing address: Institut des Sciences du Végétal, CNRS, Avenue de la Terrasse, Bât. 23, 91198 Gif sur Yvette Cedex, France. Phone: (33) 169823571. Fax: (33) 169823695. E-mail for B. C. Ramírez: ramirez{at}isv.cnrs-gif.fr. E-mail for B. Gronenborn: gronenborn{at}isv.cnrs-gif.fr.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org/.

Journal of Virology, July 2005, p. 8422-8430, Vol. 79, No. 13
0022-538X/05/$08.00+0     doi:10.1128/JVI.79.13.8422-8430.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.



This article has been cited by other articles:

  • Lageix, S., Catrice, O., Deragon, J.-M., Gronenborn, B., Pelissier, T., Ramirez, B. C. (2007). The Nanovirus-Encoded Clink Protein Affects Plant Cell Cycle Regulation through Interaction with the Retinoblastoma-Related Protein. J. Virol. 81: 4177-4185 [Abstract] [Full Text]  
  • Timchenko, T., Katul, L., Aronson, M., Vega-Arreguin, J. C., Ramirez, B. C., Vetten, H. J., Gronenborn, B. (2006). Infectivity of nanovirus DNAs: induction of disease by cloned genome components of Faba bean necrotic yellows virus. J. Gen. Virol. 87: 1735-1743 [Abstract] [Full Text]  


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