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Journal of Virology, July 2004, p. 7698-7706, Vol. 78, No. 14
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.14.7698-7706.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Interaction of DNA with the Movement Proteins of Geminiviruses Revisited

Stefan Hehnle, Christina Wege, and Holger Jeske*

Department of Molecular Biology and Plant Virology, Institute of Biology, University of Stuttgart, D-70550 Stuttgart, Germany

Received 26 December 2003/ Accepted 15 March 2004

Geminiviruses manage the transport of their DNA within plants with the help of three proteins, the coat protein (CP), the nuclear shuttle protein (NSP), and the movement protein (MP). The DNA-binding capabilities of CP, NSP, and MP of Abutilon mosaic virus (AbMV; family Geminiviridae; genus Begomovirus) were scrutinized using gel mobility shift assays and electron microscopy. CP and NSP revealed a sequence-independent affinity for both double-stranded and single-stranded DNA, as has been previously reported for other begomoviruses. MP interacted selectively with dimeric supercoiled plasmid DNA in the electrophoretic assay. Further apparent size- and form-selective binding capacities of MP have been previously reported for another geminivirus (Bean dwarf mosaic virus), but in the case of AbMV, they have been identified as the result of electrophoretic interference rather than of complex formation. Without these complications, electron microscopy confirmed the assembly of double-stranded supercoiled DNA with NSP and MP into conspicuous structures and provided the first direct evidence for cooperative interaction of MP, NSP, and DNA. Based on these results and previous ones, a transport model of geminiviruses is discussed in which NSP packages DNA and MP anchors this complex to the protoplasmic leaflets of plasma membranes and microsomes for cell-to-cell movement.

* Corresponding author. Mailing address: Department of Molecular Biology and Plant Virology, Institute of Biology, University of Stuttgart, Pfaffenwaldring 57, D-70550 Stuttgart, Germany. Phone: 49-711-685-5070. Fax: 49-711-85-5096. E-mail: holger.jeske{at}po.uni-stuttgart.de.

Journal of Virology, July 2004, p. 7698-7706, Vol. 78, No. 14
0022-538X/04/$08.00+0     DOI: 10.1128/JVI.78.14.7698-7706.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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