Human Cytomegalovirus Influences Host circRNA Transcriptions during Productive Infection
Human Cytomegalovirus Influences Host circRNA Transcriptions during Productive Infection
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人类巨细胞病毒在生产性感染期间影响宿主 circRNA 转录
DOI：
10.1007/s12250-020-00275-6
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发表时间：
2020-08-05
影响因子：
5.5
通讯作者：
Ruan, Qiang
中科院分区：
文献类型：
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作者：
Deng, Jingui;Huang, Yujing;Ruan, Qiang
人类巨细胞病毒（HCMV）是一种广泛感染人类的双链DNA病毒。环状RNA（circRNA）是非编码RNA，其大部分功能尚不清楚，并且HCMV产生性感染对宿主circRNA转录的影响仍不清楚。在这项研究中，我们通过 RNA 深度测序和生物信息学分析，分析了人胚胎肺成纤维细胞 (HELF) 中 283 个因 HCMV 生产性感染而显着改变的宿主 circRNA。其中，circSP100、circMAP3K1、circPLEKHM1 和 circTRIO 的转录和序列经过验证。此外，通过 RT-qPCR 和 Northern 印迹研究了 circSP100 的特征。这暗示circSP100是由含有六个外显子的SP100基因的有义链产生的。感染后circSP100和SP100 mRNA的动力学存在显着差异：circSP100水平随着感染而逐渐增加，而SP100 mRNA水平在感染后24小时（hpi）开始增加并下降。同时，通过RNA反义纯化（RAP）和质谱分析，总共鉴定出257个蛋白质，包括10个HCMV编码蛋白质，可能与细胞质circSP100结合。富集分析显示这些蛋白主要参与剪接体、蛋白加工、核糖体和吞噬体途径，表明 circSP100 在 HCMV 感染过程中具有多种功能。
Human cytomegalovirus (HCMV) is a double-strand DNA virus widely infected in human. Circular RNAs (circRNAs) are non-coding RNAs with most functions of which keep unknown, and the effects of HCMV productive infection on host circRNA transcriptions remain unclear. In this study, we profiled 283 host circRNAs that significantly altered by HCMV productive infection in human embryonic lung fibroblasts (HELF) by RNA deep sequencing and bioinformatics analysis. Among these, circSP100, circMAP3K1, circPLEKHM1, and circTRIO were validated for their transcriptions and sequences. Furthermore, characteristics of circSP100 were investigated by RT-qPCR and northern blot. It was implied that circSP100 was produced from the sense strand of the SP100 gene containing six exons. Kinetics of circSP100 and SP100 mRNA were significantly different after infection: circSP100 levels increased gradually along with infection, whereas SP100 mRNA levels increased in the beginning and dropped at 24 h post-infection (hpi). Meanwhile, a total number of 257 proteins, including 10 HCMV encoding proteins, were identified potentially binding to cytoplasmic circSP100 by RNA antisense purification (RAP) and mass spectrometry. Enrichment analysis showed these proteins were mainly involved in the spliceosome, protein processing, ribosome, and phagosome pathways, suggesting multiple functions of circSP100 during HCMV infection.