Self-interaction of Abutilon mosaic virus replication initiator protein (Rep) in plant cell nuclei |
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Authors: | Krenz Björn Neugart Felix Kleinow Tatjana Jeske Holger |
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Affiliation: | aInstitute of Biology, Dpt. of Molecular Biology and Plant Virology, Pfaffenwaldring 57, University of Stuttgart, D-70550 Stuttgart, Germany;bInstitute of Cell Biology, Allmandring 30, University of Stuttgart, D-70550 Stuttgart, Germany |
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Abstract: | Geminiviruses replicate their circular single-stranded DNA genome in nuclei of infected plant cells. Their replication initiator proteins (Reps) possess interaction domains for homo- and hetero-oligomerization as shown previously by in vitro studies and yeast two hybrid assays. Here, homo-oligomerization and cellular localization of the Abutilon mosaic virus (AbMV) Rep was analysed with bimolecular fluorescence complementation (BiFC) in epidermal tissues of Nicotiana benthamiana. BiFC revealed that Rep oligomers accumulated within the nucleoplasm, but were excluded from nucleoli as indicated by a nucleoli/cajal body marker. A similar subcellular distribution was observed for Rep fused to full-length cyan fluorescent protein. To examine whether tagged Reps were functionally active, N. benthamiana plants transgenic for a dimeric AbMV DNA B were inoculated with the BiFC expression constructs and nucleic acids were analysed by rolling circle amplification/restriction fragment length polymorphism as well as Southern blot hybridization. The results confirmed that the modified AbMV Rep was able to transreplicate DNA B. |
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Keywords: | Geminivirus Begomovirus Replication |
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