Rotavirus mRNAS are released by transcript-specific channels in the double-layered viral capsid
Rotavirus mRNAS are released by transcript-specific channels in the double-layered viral capsid
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DOI：
10.1073/pnas.1220345110
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发表时间：
2013-07-16
影响因子：
11.1
通讯作者：
Kapanidis, Achillefs N.
中科院分区：
文献类型：
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作者：
Periz, Javier;Celma, Cristina;Kapanidis, Achillefs N.
轮状病毒是全世界致命和严重儿童腹泻疾病的单一最常见原因（每年有1.25亿例）。轮状病毒与许多病毒具有相同的结构和功能特征，例如选择性地存在分段双链RNA基因组，并在二十面体衣壳中紧密包裹着保守数量的转录复合物。新生转录本通过12个通道离开衣壳，但尚不清楚这些通道是专门针对特定转录本还是仅仅作为一般的出口通道；了解病毒复制和基因组组织需要详细描述这一过程。为此，我们开发了一种单分子检测方法，用于捕获和鉴定从转录活性病毒颗粒中挤出的转录本。我们的研究结果支持一个模型，其中每个通道专门挤出一个特定片段的转录本，而这个转录本又与一个单一的转录复合体相连。我们的方法可以扩展到研究其他病毒和转录系统。
Rotaviruses are the single most common cause of fatal and severe childhood diarrheal illness worldwide (>125 million cases annually). Rotavirus shares structural and functional features with many viruses, such as the presence of segmented double-stranded RNA genomes selectively and tightly packed with a conserved number of transcription complexes in icosahedral capsids. Nascent transcripts exit the capsid through 12 channels, but it is unknown whether these channels specialize in specific transcripts or simply act as general exit conduits; a detailed description of this process is needed for understanding viral replication and genomic organization. To this end, we developed a single molecule assay for capturing and identifying transcripts extruded from transcriptionally active viral particles. Our findings support a model in which each channel specializes in extruding transcripts of a specific segment that in turn is linked to a single transcription complex. Our approach can be extended to study other viruses and transcription systems.