Template RNA Length Determines the Size of Replication Complex Spherules for Semliki Forest Virus
Template RNA Length Determines the Size of Replication Complex Spherules for Semliki Forest Virus
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DOI：
10.1128/jvi.00660-13
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发表时间：
2013-08-01
影响因子：
5.4
通讯作者：
Ahola, Tero
中科院分区：
文献类型：
--
作者：
Kallio, Katri;Hellstrom, Kirsi;Ahola, Tero
正链RNA病毒的复制复合物总是与细胞膜结合。复制相关膜的形态在不同的病毒系统中以不同的方式改变，但许多病毒诱导小的膜内陷，称为小球作为其复制位点。我们在这里表明，对于Semliki森林病毒（SFV），一种甲病毒，小球的大小与复制RNA模板的长度密切相关。用相关的光学和电子显微镜分析了具有不同模型模板的细胞，其通过病毒复制酶反式表达和复制。结果表明，11.5 kb的病毒基因组大小的模板诱导的小球直径与58 nm相似，而6 kb的模板产生的小球直径与39 nm相似。通过放射性标记和北方印迹评估，不同大小的病毒模板被有效地反式复制。复制两种不同的模板，顺式和反式，在同一个细胞中产生两个大小类的小球。这些结果表明，RNA在SFV的小球组装中起着至关重要的决定作用，与其他正链RNA病毒的结果形成直接对比，其中病毒RNA的存在或RNA大小对小球形成没有贡献。
The replication complexes of positive-strand RNA viruses are always associated with cellular membranes. The morphology of the replication-associated membranes is altered in different ways in different viral systems, but many viruses induce small membrane invaginations known as spherules as their replication sites. We show here that for Semliki Forest virus (SFV), an alphavirus, the size of the spherules is tightly connected with the length of the replicating RNA template. Cells with different model templates, expressed in trans and copied by the viral replicase, were analyzed with correlative light and electron microscopy. It was demonstrated that the viral-genome-sized template of 11.5 kb induced spherules that were similar to 58 nm in diameter, whereas a template of 6 kb yielded similar to 39-nm spherules. Different sizes of viral templates were replicated efficiently in trans, as assessed by radioactive labeling and Northern blotting. The replication of two different templates, in cis and trans, yielded two size classes of spherules in the same cell. These results indicate that RNA plays a crucial determining role in spherule assembly for SFV, in direct contrast with results from other positive-strand RNA viruses, in which either the presence of viral RNA or the RNA size do not contribute to spherule formation.