Institute of Oceanography, Minjiang University, Fuzhou 350108, China.
College of Biological Sciences and Engineering, Fuzhou University, Fuzhou 350108, China.
J Invertebr Pathol. 2022 Jan;187:107699. doi: 10.1016/j.jip.2021.107699. Epub 2021 Nov 26.
Iridovirids are a group icosahedral viruses containing linear double-stranded DNA, and mainly infect invertebrates and poikilothermic vertebrates. Cherax quadricarinatus iridovirus (CQIV) is a new species of the family Iridoviridae and can cause high mortality in shrimps. In CQIV genome, there are 25 conserved genes and the putative products are involved in several viral processes. In this study, three core protein including CQIV-032R, CQIV-125R and CQIV-160L were identified to interact with CQIV-038R by yeast two-hybrid (Y2H), and the interaction between CQIV-038R and CQIV-125R was further confirmed by co-immunoprecipitation (Co-IP) assays. In the expression system, EGFP-038R and mCherry-125R were colocalized in the cytoplasm when co-expressed in Sf9 cells. Moreover, silencing the expression of 038R, 125R or both of these two proteins respectively in C. quadricarinatus cells by small interfering RNAs showed significantly inhibit CQIV replication. Collectively, we identified the interaction between 038R and 125R, and demonstrated they are essential for CQIV replication.
虹彩病毒是一类含有线性双链 DNA 的二十面体病毒，主要感染无脊椎动物和变温脊椎动物。克氏原螯虾虹彩病毒（Cherax quadricarinatus iridovirus，CQIV）是虹彩病毒科的一个新种，可引起虾类的高死亡率。在 CQIV 基因组中，有 25 个保守基因，其假定产物参与了几种病毒过程。在本研究中，通过酵母双杂交（yeast two-hybrid，Y2H）鉴定出三个核心蛋白，包括 CQIV-032R、CQIV-125R 和 CQIV-160L，它们与 CQIV-038R 相互作用，并且通过共免疫沉淀（co-immunoprecipitation，Co-IP）实验进一步证实了 CQIV-038R 和 CQIV-125R 之间的相互作用。在表达系统中，当在 Sf9 细胞中共表达时，EGFP-038R 和 mCherry-125R 共定位于细胞质中。此外，通过小干扰 RNA 分别沉默 C. quadricarinatus 细胞中 038R、125R 或这两种蛋白的表达，显著抑制了 CQIV 的复制。综上所述，我们鉴定了 038R 和 125R 之间的相互作用，并证明它们是 CQIV 复制所必需的。
