Activities associated with the putative replication initiation protein of Coconut foliar decay virus, a tentative member of the genus Nanovirus

• Published in: Journal of general virology Vol. 81; no. 12; pp. 3099 - 3106
• Main Authors: Merits, Andres, Fedorkin, Oleg N, Guo, Deyin, Kalinina, Natalia O, Morozov, Sergey Yu
• Format: Journal Article
• Language: English
• Published: England Soc General Microbiol 01.12.2000
• Subjects: Adenosine Triphosphatases - genetics; Adenosine Triphosphatases - metabolism; Animals; Cell Line; Cell Membrane - metabolism; Cell Membrane - virology; Cell Nucleus - metabolism; Cell Nucleus - virology; Coconut foliar decay virus; Cocos - virology; DNA - metabolism; DNA Helicases - genetics; DNA Helicases - metabolism; DNA Replication; DNA Viruses - enzymology; DNA Viruses - genetics; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Escherichia coli; Geminiviridae - enzymology; GTP Phosphohydrolases - genetics; GTP Phosphohydrolases - metabolism; Microscopy, Fluorescence; Nicotiana; Nicotiana benthamiana; Plant Viruses - enzymology; Plant Viruses - genetics; Plants, Toxic; Protein Binding; Recombinant Fusion Proteins - genetics; Recombinant Fusion Proteins - metabolism; RNA - metabolism; RNA-Binding Proteins - genetics; RNA-Binding Proteins - metabolism; Spodoptera; Trans-Activators - genetics; Trans-Activators - metabolism; Two-Hybrid System Techniques; Viral Proteins - genetics; Viral Proteins - metabolism
• ISSN: 0022-1317; 1465-2099
• DOI: 10.1099/0022-1317-81-12-3099

National Institute of Chemical Physics and Biophysics, Akadeemia tee 23, EE12618 Tallinn, Estonia 1 Institute of Biotechnology, Viikki Biocenter, University of Helsinki, PO Box 56, FIN-00014 Helsinki, Finland 2 Department of Virology and A. N. Belozersky Institute of Physico-Chemical Biology, Moscow State University, Moscow 119899, Russia 3 Author for correspondence: Andres Merits (at the Viikki Biocenter). Fax +358 9 191959366. e-mail Merits{at}operoni.helsinki.fi The putative replication initiation protein (Rep) of Coconut foliar decay virus (CFDV) was expressed as a 6 x His recombinant protein in E. coli and in recombinant baculovirus. Purified 6 x His–Rep protein was demonstrated to possess sequence non-specific RNA- and ssDNA-binding activities as well as magnesium-dependent ATPase/GTPase activity. The yeast two-hybrid system revealed that CFDV Rep could interact with itself. Subcellular distribution of the CFDV Rep was studied by fractionation of insect cells infected with recombinant baculovirus expressing the 6 x His–Rep protein and by laser scanning confocal microscopy of Nicotiana benthamiana epidermal cells bombarded with a construct encoding CFDV Rep fused to GFP. It was shown that CFDV Rep associated predominantly with nuclei and membranes of infected/transfected cells. These activities of CFDV-encoded Rep are very similar to those reported for Reps of geminiviruses.