Viruses 2013, 5(10), 2512-2530; doi:10.3390/v5102512
In planta Protein Interactions of Three Alphacryptoviruses and Three Betacryptoviruses from White Clover, Red Clover and Dill by Bimolecular Fluorescence Complementation Analysis
Department of Phytomedicine, Institute of Horticultural Production Systems, Leibniz Universität Hannover, Herrenhäuser Str. 2, Hannover D-30419, Germany
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Received: 20 August 2013 / Revised: 25 September 2013 / Accepted: 27 September 2013 / Published: 9 October 2013
(This article belongs to the Section Viruses of Plants, Fungi and Protozoa)
Abstract
Plant-infecting viruses of the genera Alpha- and Betacryptovirus within the family Partitiviridae cause no visible effects on their hosts and are only transmitted by cell division and through gametes. The bipartite dsRNA genome is encoding a RNA-dependent RNA polymerase (RdRp) and a coat protein (CP). Aside from sequence and structural analysis, the investigation of protein interactions is another step towards virus characterization. Therefore, ORFs of two type members White Clover Cryptic Virus 1 and 2 (WCCV-1 and WCCV-2), as well as the related viruses from Red Clover and Dill were introduced into a bimolecular fluorescence complementation assay. We showed CP-CP dimerization for all tested viruses with localization for alphacryptoviruses at the nuclear membrane and for betacryptoviruses close to cell walls within the cytoplasm. For CPs of WCCV-1 and WCCV-2, deletion mutants were created to determine internal interaction sites. Moreover, RdRp self-interaction was found for all viruses, whereas CP-RdRp interactions were only detectable for the alphacryptoviruses. An intra-genus test of CPs was successful in various virus combinations, whereas an inter-genus interaction of WCCV-1CP and WCCV-2CP was absent. This is the first report of in vivo protein interactions of members in the family Partitiviridae, indicating distinct features of the alpha- and betacryptoviruses. View Full-TextKeywords:
Partitiviridae; Alphacryptovirus; Betacryptovirus; protein interaction; Bimolecular fluorescence complementation (BiFC)
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Lesker, T.; Maiss, E. In planta Protein Interactions of Three Alphacryptoviruses and Three Betacryptoviruses from White Clover, Red Clover and Dill by Bimolecular Fluorescence Complementation Analysis. Viruses 2013, 5, 2512-2530.