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Viruses 2013, 5(10), 2512-2530; doi:10.3390/v5102512
Article

In planta Protein Interactions of Three Alphacryptoviruses and Three Betacryptoviruses from White Clover, Red Clover and Dill by Bimolecular Fluorescence Complementation Analysis

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Received: 20 August 2013; in revised form: 25 September 2013 / Accepted: 27 September 2013 / Published: 9 October 2013
(This article belongs to the Section Viruses of Plants, Fungi and Protoza)
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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.
Keywords: Partitiviridae; Alphacryptovirus; Betacryptovirus; protein interaction; Bimolecular fluorescence complementation (BiFC) Partitiviridae; Alphacryptovirus; Betacryptovirus; protein interaction; Bimolecular fluorescence complementation (BiFC)
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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MDPI and ACS Style

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.

AMA Style

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(10):2512-2530.

Chicago/Turabian Style

Lesker, Till; Maiss, Edgar. 2013. "In planta Protein Interactions of Three Alphacryptoviruses and Three Betacryptoviruses from White Clover, Red Clover and Dill by Bimolecular Fluorescence Complementation Analysis." Viruses 5, no. 10: 2512-2530.



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