Deep Sequencing Reveals Direct Targets of Gammaherpesvirus-Induced mRNA Decay and Suggests That Multiple Mechanisms Govern Cellular Transcript Escape
Deep Sequencing Reveals Direct Targets of Gammaherpesvirus-Induced mRNA Decay and Suggests That Multiple Mechanisms Govern Cellular Transcript Escape
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DOI：
10.1371/journal.pone.0019655
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发表时间：
2011-05-09
期刊：
影响因子：
3.7
通讯作者：
Glaunsinger, Britt A.
中科院分区：
文献类型：
--
作者：
Clyde, Karen;Glaunsinger, Britt A.
包括卡波西肉瘤相关疱疹病毒（KSHV）、EB病毒（EBV）和鼠疱疹病毒68（MHV 68）在内的γ疱疹病毒裂解感染的一个特征是在称为宿主关闭的过程中显著抑制细胞基因表达。碱性核酸外切酶蛋白（KSHV SOX、MHV-68 muSOX和EBV BGLF 5）已显示通过使细胞mRNA不稳定来诱导关闭。在这里，我们扩展了先前对裂解感染期间细胞mRNA丰度的分析，以在不存在其他病毒基因的情况下，利用深度测序技术（RNA-seq）来表征SOX和muSOX的影响。与先前在裂解性感染期间的观察一致，大多数转录物在表达SOX或muSOX的细胞中下调，其中muSOX充当比SOX更有效的关闭因子。此外，大多数细胞信息在SOX和muSOX表达细胞中属于相同的表达类别，表明两种因子靶向相似的mRNA库。更丰富的mRNA被更有效地下调，表明转录靶向中的浓度效应。然而，即使在高表达的基因中，也有一些mRNA能够逃脱宿主的关闭。选择逃逸的进一步表征揭示了细胞基因可以逃避下调的多种机制。虽然一些mRNA对SOX直接不敏感，但其他mRNA的稳态水平保持不变，推测是mRNA生物合成的下游效应的结果。总的来说，这些研究奠定了框架解剖的机制，潜在的敏感性的mRNA破坏裂解γ疱疹病毒感染。
One characteristic of lytic infection with gammaherpesviruses, including Kaposi's sarcoma-associated herpesvirus (KSHV), Epstein-Barr virus (EBV) and murine herpesvirus 68 (MHV68), is the dramatic suppression of cellular gene expression in a process known as host shutoff. The alkaline exonuclease proteins (KSHV SOX, MHV-68 muSOX and EBV BGLF5) have been shown to induce shutoff by destabilizing cellular mRNAs. Here we extend previous analyses of cellular mRNA abundance during lytic infection to characterize the effects of SOX and muSOX, in the absence of other viral genes, utilizing deep sequencing technology (RNA-seq). Consistent with previous observations during lytic infection, the majority of transcripts are downregulated in cells expressing either SOX or muSOX, with muSOX acting as a more potent shutoff factor than SOX. Moreover, most cellular messages fall into the same expression class in both SOX- and muSOX-expressing cells, indicating that both factors target similar pools of mRNAs. More abundant mRNAs are more efficiently downregulated, suggesting a concentration effect in transcript targeting. However, even among highly expressed genes there are mRNAs that escape host shutoff. Further characterization of select escapees reveals multiple mechanisms by which cellular genes can evade downregulation. While some mRNAs are directly refractory to SOX, the steady state levels of others remain unchanged, presumably as a consequence of downstream effects on mRNA biogenesis. Collectively, these studies lay the framework for dissecting the mechanisms underlying the susceptibility of mRNA to destruction during lytic gammaherpesvirus infection.