Abstract
Southern rice black streaked dwarf virus (SRBSDV) causes severe harm to rice production. Unfortunately, studies on effective antiviral drugs against SRBSDV and interaction mechanism of antiviral molecule targeting on SRBSDV have not been reported. This study found dufulin (DFL), an ideal anti-SRBSDV molecule, and investigated the interactions of DFL targeting on the nonstructural protein P9-1. The biological sequence information and bonding characterization of DFL to four kinds of P9-1 protein were described with fluorescence titration (FT) and microscale thermophoresis (MST) assays. The sequence analysis indicated that P9-1 had highly-conserved C- and N-terminal amino acid residues and a hypervariable region that differed from 131 aa to 160 aa. Consequently, wild-type (WT-His-P9-1), 23 C-terminal residues truncated (TR-ΔC23-His-P9-1), 6 N-terminal residues truncated (TR-ΔN6-His-P9-1), and Ser138 site-directed (MU-138-His-P9-1) mutant proteins were expressed. The FT and MST assay results indicated that DFL bounded to WT-His-P9-1 with micromole affinity and the 23 C-terminal amino acids were the potential targeting site. This system, which combines a complete sequence analysis, mutant protein expression, and binding action evaluating system, could further advance the understanding of the interaction abilities between antiviral drugs and their targets.
Highlights
Plant-infecting reoviruses are classified into three genera, namely Phytoreovirus, Oryzavirus and Fijivirus
According to the results of the nine complete nucleotide sequences, Southern rice black streaked dwarf virus (SRBSDV)-P9 consisted of 1900 base pairs, similar to other reported SRBSDV isolates downloaded from NCBI
The alignment results indicated that P9-1 protein had highly conserved C- and N-terminal amino acid residues and a hypervariable region that differ from 131 aa to 160 aa compared with rice black streaked dwarf virus (RBSDV) (Figure 2)
Summary
Plant-infecting reoviruses are classified into three genera, namely Phytoreovirus, Oryzavirus and Fijivirus. Fijivirus species containing 10 segments, with an aggregate genome size larger than in other plant reovirus genera are distributed worldwide, and some of them cause serious diseases [1,2]. Southern rice black streaked dwarf virus (SRBSDV) transmitted by white-backed planthopper (Sogatella furcifera Horváth) vectors in a persistent propagative manner is a member of the genus Fijivirus of the Reoviridae family; it is a non-enveloped double-stranded RNA virus that has caused significant losses of grain yields in parts of Asia, including Vietnam, Japan, and China [3,4,5,6,7]. A comparison of the different genomic segments of SRBSDV with those of their counterparts in rice black streaked dwarf virus (RBSDV) suggests that SRBSDV encodes at least six putative structural proteins (P1, P2, P3, P4, P8, and P10) and five putative nonstructural proteins (P6, P7-1, P7-2, P9-1, and P9-2) [9]. Among the putative nonstructural proteins encoded by SRBSDV, P6 is a viral RNA-silencing suppressor [14], and P7-1 has the intrinsic ability to self-interact to form tubules in non-host insect cells [15]
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