Identification of infectious spleen and kidney necrosis virus (ISKNV)-encoded microRNAs
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ORIGINAL PAPER
Identification of infectious spleen and kidney necrosis virus (ISKNV)‑encoded microRNAs Jian‑hui He1 · Qiong Xia1 · Shaoping Weng1,2,3 · Jianguo He1,2,3 · Xiaopeng Xu1,2,3 Received: 5 May 2020 / Accepted: 25 September 2020 © Springer Science+Business Media, LLC, part of Springer Nature 2020
Abstract MicroRNAs (miRNAs) are small noncoding RNAs that post-transcriptionally regulate gene expression by complementary binding to target mRNAs. Virus-encoded miRNAs play important roles in virus life cycle and virus-host interactions. Viruses from the Megalocytivirus genus, family Iridoviridae, infect a wide range of fishes, bringing great challenges to aquaculture. Infectious spleen and kidney necrosis virus (ISKNV) is the type species of the Megalocytivirus genus. In this study, using Illumina sequencing coupled with miRNA precursor prediction and stem-loop real-time PCR, 14 putative ISKNV-encoded miRNAs were preliminarily identified from ISKNV-infected mandarin fish MFF-1 cells. To initially study their functions, inhibitors of the 14 viral miRNAs were synthesized and transfected into MFF-1 cells, which were further infected with ISKNV. The results showed that these viral miRNAs could affect the virus titers in the supernatant of ISKNV-infected cells and the expression of major capsid protein (MCP). Moreover, we observed that inhibition of several ISKNV miRNAs had different effects on MCP expression and on titer of released virus, suggesting complex roles of viral miRNAs in ISKNV infection. The current study may provide a fundamental information for further identification and functional studies on miRNAs encoded by Megalocytivirus.
Introduction microRNAs (miRNAs) are a group of small noncoding RNAs (~ 22 nt) that functionally participate in various biological processes by post-transcriptionally regulating gene expression [1, 2]. miRNA genes are initially transcribed into primary miRNAs (pri-miRNAs, ~ 1000 nt), which are further processed into stem-loop-shaped precursor miRNAs (premiRNAs, 70 ~ 100 nt) [3]. After exported from the nucleus, Edited by Juergen Richt. * Jianguo He [email protected] * Xiaopeng Xu [email protected] 1
State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, People’s Republic of China
2
Southern Marine Science and Engineering Guangdong Laboratory (Zhuhai), Sun Yat-Sen University, Guangzhou, People’s Republic of China
3
Institute of Aquatic Economic Animals and Guangdong Provice Key Laboratory for Aquatic Economic Animals, Sun Yat-Sen University, Guangzhou, People’s Republic of China
pre-miRNAs are further processed into mature miRNAs by the Dicer complex [4]. In an ATP-dependent manner, mature miRNAs are incorporated into a RNA-induced silencer complex (RISC)-loading complex (RLC) and act on the 3′ UTR or open reading frames (ORFs) of target mRNAs, resulting in gene expression silencing [5, 6]. Since virus-encoded miRNAs were firstly identified in Epstein–Barr virus (EBV) in 2004 [7], more and more viral mi
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