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Characterization of Striped jack nervous necrosis virus subgenomic RNA3 and biological activities of its encoded protein B2

亚基因组mRNA 生物 病毒学 核糖核酸 分子生物学 病毒 RNA病毒 RNA沉默 基因 RNA干扰 遗传学
作者
Tokinori Iwamoto,Kazuyuki Mise,Atsushi Takeda,Yasushi Okinaka,Koh-ichiro Mori,Misao Arimoto,Tetsuro Okuno,Toshihiro Nakai
出处
期刊:Journal of General Virology [Microbiology Society]
卷期号:86 (10): 2807-2816 被引量:166
标识
DOI:10.1099/vir.0.80902-0
摘要

Striped jack nervous necrosis virus (SJNNV), which infects fish, is the type species of the genus Betanodavirus. This virus has a bipartite genome of positive-strand RNAs, designated RNAs 1 and 2. A small RNA (ca. 0.4 kb) has been detected from SJNNV-infected cells, which was newly synthesized and corresponded to the 3'-terminal region of RNA1. Rapid amplification of cDNA ends analysis showed that the 5' end of this small RNA (designated RNA3) initiated at nt 2730 of the corresponding RNA1 sequence and contained a 5' cap structure. Substitution of the first nucleotide of the subgenomic RNA sequence within RNA1 selectively inhibited production of the positive-strand RNA3 but not of the negative-strand RNA3, which suggests that RNA3 may be synthesized via a premature termination model. The single RNA3-encoded protein (designated protein B2) was expressed in Escherichia coli, purified and used to immunize a rabbit to obtain an anti-protein B2 polyclonal antibody. An immunological test showed that the antigen was specifically detected in the central nervous system and retina of infected striped jack larvae (Pseudocaranx dentex), and in the cytoplasm of infected cultured E-11 cells. These results indicate that SJNNV produces subgenomic RNA3 from RNA1 and synthesizes protein B2 during virus multiplication, as reported for alphanodaviruses. In addition, an Agrobacterium co-infiltration assay established in transgenic plants that express green fluorescent protein showed that SJNNV protein B2 has a potent RNA silencing-suppression activity, as discovered for the protein B2 of insect-infecting alphanodaviruses.

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