长链非编码RNA CTO-S对伪狂犬病毒复制的影响

Effect of long non-coding RNA CTO-S on the replication of pseudorabies virus

摘要:

【目的】研究伪狂犬病毒（pseudorabies virus，PRV）的长链非编码RNA （long noncoding RNA，lncRNA） CTO-S对于病毒复制的影响。【方法】利用3'RACE和5'RACE鉴定CTO-S的全长序列。验证CTO-S是否具有编码多肽的能力。Red重组方法构建CTO-S缺失株以及回复突变株。分析野毒株、缺失株和回复突变株复制的差异。利用RNA pull down联合质谱的方法筛选与CTO-S互作的宿主蛋白。利用RNA免疫共沉淀（RNA immunoprecipitation，RIP）方法进行验证。最后利用四甲基偶氮唑盐（MTT）法检测野毒株、缺失株和回复突变株对细胞凋亡的影响。【结果】CTO-S全长267 bp，不具有编码多肽的能力。CTO-S缺失不影响PRV的复制。CTO-S与泛醇细胞色素c还原酶的组成部分cytochrome b-c1 complex subunit 1互作，诱导细胞凋亡。【结论】CTO-S对于PRV的复制是非必需的，CTO-S与泛醇细胞色素c还原酶的组成部分cytochrome b-c1 complex subunit 1互作，诱导细胞凋亡。

Abstract:

[Objective] This study aims to reveal the effect of the long non-coding RNA (lncRNA) CTO-S on the replication of pseudorabies virus (PRV). [Methods] We employed 3'RACE and 5'RACE to identify the full-length sequence of CTO-S, and verified whether CTO-S had the ability to encode polypeptides. After constructing the CTO-S deletion and complementation strains with the Red recombination method, we analyzed the differences in the replication of wild-type, deletion, and complementation strains. We used RNA pull down combined with mass spectrometry to screen out the host protein interacting with CTO-S, and then adopted RNA immunoprecipitation (RIP) method to verify the interaction between the screened protein and CTO-S. Finally, the methyl tetrazolium (MTT) method was used to detect the effects of wild-type, deletion, and complementation strains on cell apoptosis. [Results] CTO-S had a full length of 267 bp and did not have the ability to encode polypeptides. The deletion of CTO-S did not affect the replication of PRV. CTO-S interacted with cytochrome b-c1 complex subunit 1, a component of ubiquinol-cytochrome c reductase, and induced apoptosis. [Conclusion] CTO-S is not essential for the replication of PRV. It interacts with cytochrome b-c1 complex subunit 1, a component of ubiquinol-cytochrome c reductase, and induces cell apoptosis.