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Article Dans Une Revue Journal of Virology Année : 2005

A functional histidine-tagged replication initiator protein: implications for the study of single-stranded DNA virus replication in planta.

Résumé

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.

Dates et versions

hal-00119941 , version 1 (12-12-2006)

Identifiants

Citer

Julio C Vega-Arreguín, Tatiana Timchenko, Bruno Gronenborn, Bertha Cecilia Ramírez. A functional histidine-tagged replication initiator protein: implications for the study of single-stranded DNA virus replication in planta.. Journal of Virology, 2005, 79 (13), pp.8422-30. ⟨10.1128/JVI.79.13.8422-8430.2005⟩. ⟨hal-00119941⟩
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