Abstract
BackgroundRice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus within the family Reoviridae, causes severe damage to cereal crops in South East Asia. The protein P7-2, encoded by the second open reading frame of segment S7, is conserved among most plant-infecting fijiviruses, but its function is still obscure.ResultsIn this study, P7-2 was used as bait in two-hybrid screens of a cDNA library expressing Zea mays proteins. It was found that there is a strong interaction between P7-2 and Z. mays SKP1 (SKP1Maize), a core subunit of the multicomponent SCF (SKP1/Cullin1/F-box/Rbx1) E3 ubiquitin ligase. The interaction was then confirmed in leaf epidermal cells of Nicotiana benthamiana by bimolecular fluorescence complementation assay. Further investigations indicated that P7-2 also interacts with SKP1 proteins from other plants, including Arabidopsis thaliana, N. benthamiana,Oryza sativa and Saccharum sinense. The C-terminal fragment of SKP1Maize (residues 97–176) and the middle fragment of P7-2 (residues 79–214) are necessary to sustain the interaction, while the C-terminal putative α-helix domain spanning residues 214–295 of P7-2 greatly facilitates the interaction. Agrobacterium-mediated transient suppression assay showed that P7-2 has no obvious activity to suppress local RNA silencing.ConclusionsTaken together, our results indicated that RBSDV P7-2 can interact with SKP1 proteins from different plants. This is the first report linking a Fijivirus protein to a component of the ubiquitin proteasome system. P7-2 might be a potential F-box protein encoded by RBSDV and involved in the plant-virus interaction through ubiquitination pathway.
Highlights
Rice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus within the family Reoviridae, causes severe damage to cereal crops in South East Asia
P7-2 interacts with SKP1 from Z. mays in yeast The potential functions of P7-2 in virus multiplication and viral pathogenicity on plants might be due to some unrevealed interactions between P7-2 and plant hosts
Making sure that the fused protein can be expressed in yeast and has no transcriptional activation or toxicity, BD-P7-2 was used as bait in two-hybrid screens of a cDNA library expressing Z. mays proteins fused to the GAL4 activation domain (AD)
Summary
Rice black-streaked dwarf virus (RBSDV), a member of the genus Fijivirus within the family Reoviridae, causes severe damage to cereal crops in South East Asia. The protein P7-2, encoded by the second open reading frame of segment S7, is conserved among most plant-infecting fijiviruses, but its function is still obscure. Rice black-streaked dwarf virus (RBSDV) is an insect- and plant-infecting agent that belongs to the genus Fijivirus in the family Reoviridae. P10 is the main component of the outer shell of the viral particle and can self-interact to form trimers in solution [5,10]. Both S7 and S9 have two non-overlapping ORFs and encode two nonstructural proteins. The functions of the remaining proteins are still largely unknown
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