In-Depth Analysis of Kaposi's Sarcoma-Associated Herpesvirus MicroRNA Expression Provides Insights into the Mammalian MicroRNA-Processing Machinery
In-Depth Analysis of Kaposi's Sarcoma-Associated Herpesvirus MicroRNA Expression Provides Insights into the Mammalian MicroRNA-Processing Machinery
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DOI：
10.1128/jvi.02013-09
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发表时间：
2010-01-15
影响因子：
5.4
通讯作者：
Cullen, Bryan R.
中科院分区：
文献类型：
--
作者：
Umbach, Jennifer L.;Cullen, Bryan R.
我们使用深度测序分析了潜伏感染卡波西肉瘤相关疱疹病毒（KSHV）的b细胞系BC-3中观察到的病毒microRNA （miRNA）表达模式。我们恢复了14.6 × 10(6)个总miRNA cDNA reads，其中92%来自KSHV。我们检测到11个KSHV miRNA以及所有11个预测miRNA*或miRNA双工中间体的客运链。先前报道的一个KSHV miRNA miR-K9被发现是突变失活的。该分析显示，11个KSHV mirna中有10个的5‘端基本不变，在3’端观察到明显更多的变异，这一结果与mirna的5'-近端区域对靶mRNA识别至关重要的建议一致。然而，一种KSHV miRNA miR-K10-3p在5'端以1个核苷酸（nt）的差异存在的两种异构体中被检测到，这两种相关的KSHV miRNA因此可能针对至少部分不同的mRNA群体。最后，我们还报道了在脊椎动物体细胞中首次检测到miRNA偏移rna （moRs）。moRs来源于直接位于成熟miRNA和miRNA*链两侧的初级miRNA （pri-miRNA）序列，被鉴定为位于9个KSHV miRNA的一侧或两侧。这些数据为哺乳动物细胞中miRNA加工模式提供了新的见解，并表明该过程在动物进化过程中高度保守。
We have used deep sequencing to analyze the pattern of viral microRNA (miRNA) expression observed in the B-cell line BC-3, which is latently infected with Kaposi's sarcoma-associated herpesvirus (KSHV). We recovered 14.6 x 10(6) total miRNA cDNA reads, of which a remarkable 92% were of KSHV origin. We detected 11 KSHV miRNAs as well as all 11 predicted miRNA* or passenger strands from the miRNA duplex intermediate. One previously reported KSHV miRNA, miR-K9, was found to be mutationally inactivated. This analysis revealed that the 5' ends of 10 of the 11 KSHV miRNAs were essentially invariant, with significantly more variation being observed at the 3' end, a result which is consistent with the proposal that the 5'-proximal region of miRNAs is critical for target mRNA recognition. However, one KSHV miRNA, miR-K10-3p, was detected in two isoforms differing by 1 nucleotide (nt) at the 5' end that were present at comparable levels, and these two related KSHV miRNAs are therefore likely to target at least partially distinct mRNA populations. Finally, we also report the first detection of miRNA offset RNAs (moRs) in vertebrate somatic cells. moRs, which derive from primary miRNA (pri-miRNA) sequences that immediately flank the mature miRNA and miRNA* strands, were identified flanking one or both sides of nine of the KSHV miRNAs. These data provide new insights into the pattern of miRNA processing in mammalian cells and indicate that this process is highly conserved during animal evolution.