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Viruses 2016, 8(7), 190; doi:10.3390/v8070190

Properties of African Cassava Mosaic Virus Capsid Protein Expressed in Fission Yeast

Department of Molecular Biology and Plant Virology, Institute of Biomaterials and Biomolecular Systems, University of Stuttgart, Pfaffenwaldring 57, D-70550 Stuttgart, Germany
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Academic Editor: Thomas Hohn
Received: 17 May 2016 / Revised: 22 June 2016 / Accepted: 29 June 2016 / Published: 8 July 2016
(This article belongs to the Section Viruses of Plants, Fungi and Protozoa)
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Abstract

The capsid proteins (CPs) of geminiviruses combine multiple functions for packaging the single-stranded viral genome, insect transmission and shuttling between the nucleus and the cytoplasm. African cassava mosaic virus (ACMV) CP was expressed in fission yeast, and purified by SDS gel electrophoresis. After tryptic digestion of this protein, mass spectrometry covered 85% of the amino acid sequence and detected three N-terminal phosphorylation sites (threonine 12, serines 25 and 62). Differential centrifugation of cell extracts separated the CP into two fractions, the supernatant and pellet. Upon isopycnic centrifugation of the supernatant, most of the CP accumulated at densities typical for free proteins, whereas the CP in the pellet fraction showed a partial binding to nucleic acids. Size-exclusion chromatography of the supernatant CP indicated high order complexes. In DNA binding assays, supernatant CP accelerated the migration of ssDNA in agarose gels, which is a first hint for particle formation. Correspondingly, CP shifted ssDNA to the expected densities of virus particles upon isopycnic centrifugation. Nevertheless, electron microscopy did not reveal any twin particles, which are characteristic for geminiviruses. View Full-Text
Keywords: geminivirus; capsid protein; CP; fission yeast; ectopic expression; DNA binding assay geminivirus; capsid protein; CP; fission yeast; ectopic expression; DNA binding assay
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. (CC BY 4.0).

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MDPI and ACS Style

Hipp, K.; Schäfer, B.; Kepp, G.; Jeske, H. Properties of African Cassava Mosaic Virus Capsid Protein Expressed in Fission Yeast. Viruses 2016, 8, 190.

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