Quantitative Analysis of MicroRNAs in Vaccinia virus Infection Reveals Diversity in Their Susceptibility to Modification and Suppression.
Quantitative Analysis of MicroRNAs in Vaccinia virus Infection Reveals Diversity in Their Susceptibility to Modification and Suppression.
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DOI：
10.1371/journal.pone.0131787
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发表时间：
2015
期刊：
影响因子：
3.7
通讯作者：
Beard PM
中科院分区：
文献类型：
--
作者：
Buck AH;Ivens A;Gordon K;Craig N;Houzelle A;Roche A;Turnbull N;Beard PM
痘苗病毒 (VACV) 是一种大型细胞质 DNA 病毒，可导致许多细胞途径发生巨大改变，包括 microRNA 生物发生。该病毒编码一种聚腺苷酸聚合酶，此前研究表明该酶可将聚腺苷酸尾添加到细胞 miRNA 的 3' 末端，导致其在感染后 24 小时 (hpi) 降解。在这里，我们使用小 RNA 测序来量化 VACV 感染在感染后早期（6 小时）和晚期（24 小时）对人类细胞中细胞 miRNA 的影响。对单个 miRNA 的详细定量分析揭示了它们在感染过程中修饰程度和丰度相对变化的显着多样性。一些 miRNA 被高度修饰（例如 miR-29a-3p、miR-27b-3p），而其他 miRNA 则出现耐药性（例如 miR-16-5p）。此外，在 6 hpi 时高度加尾的 miRNA 不一定在 24 hpi 时减少最多。这些结果表明，人类细胞 miRNA 的内在特征导致它们在 VACV 感染期间发生差异性多腺苷酸化和丰度改变。我们还证明，中间和晚期 VACV 基因表达对于包括 miR-27-3p 在内的一些 miRNA 的最佳抑制是必需的。总体而言，这项工作揭示了 VACV 感染对宿主 miRNA 的复杂多样的影响，并鉴定了对 VACV 诱导的聚腺苷酸化有很大抵抗力的 miRNA，因此在感染和复制的初始阶段以功能水平存在。
Vaccinia virus (VACV) is a large cytoplasmic DNA virus that causes dramatic alterations to many cellular pathways including microRNA biogenesis. The virus encodes a poly(A) polymerase which was previously shown to add poly(A) tails to the 3’ end of cellular miRNAs, resulting in their degradation by 24 hours post infection (hpi). Here we used small RNA sequencing to quantify the impact of VACV infection on cellular miRNAs in human cells at both early (6 h) and late (24 h) times post infection. A detailed quantitative analysis of individual miRNAs revealed marked diversity in the extent of their modification and relative change in abundance during infection. Some miRNAs became highly modified (e.g. miR-29a-3p, miR-27b-3p) whereas others appeared resistant (e.g. miR-16-5p). Furthermore, miRNAs that were highly tailed at 6 hpi were not necessarily among the most reduced at 24 hpi. These results suggest that intrinsic features of human cellular miRNAs cause them to be differentially polyadenylated and altered in abundance during VACV infection. We also demonstrate that intermediate and late VACV gene expression are required for optimal repression of some miRNAs including miR-27-3p. Overall this work reveals complex and varied consequences of VACV infection on host miRNAs and identifies miRNAs which are largely resistant to VACV-induced polyadenylation and are therefore present at functional levels during the initial stages of infection and replication.
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