2A Protease Is Not a Prerequisite for Poliovirus Replication
2A Protease Is Not a Prerequisite for Poliovirus Replication
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DOI：
10.1128/jvi.02575-09
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发表时间：
2010-06-01
影响因子：
5.4
通讯作者：
Nomoto, Akio
中科院分区：
文献类型：
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作者：
Igarashi, Hiroko;Yoshino, Yasuko;Nomoto, Akio
脊髓灰质炎病毒（PV）2A（pro）被认为对PV复制很重要，已知对宿主细胞有毒。2A（pro）缺陷型PV可能毒性较低，并且作为载体是理想的。为了检测是否需要2A（pro）来形成子代病毒，在PV 1（M）OM株中构建了具有（pOME）或不具有（pOME Delta 2A）2A（pro）的双顺反子（dc）PV的全长cDNA。pOME和pOME Delta 2A的RNA都能够形成子代病毒，分别称为OME和OME Delta 2A。在它们诱导细胞病变效应（CPE）的能力方面，被列为OME Delta 2A < OME的菌株大约等于PV 1（M）OM。这些结果表明，2A（pro）对于全长dc PV形成子代病毒不是必需的，并且其有助于有效的病毒复制和/或CPE的诱导。为了阐明2A（pro）是否是P1-null（缺乏衣壳蛋白的整个编码序列）PV所必需的，检查了P1-null PV（pOM Delta P1）或没有2A（pro）的P1-null PV（pOM Delta P1 Delta 2A）或没有2A（pro）和2B两者的P1-null PV（pOM Delta P1 Delta 2A Delta 2B）的RNA复制活性。pOM Delta P1和pOM Delta P1 Delta 2A的RNA在P1蛋白的反式供应下可以复制并形成子代病毒，而pOM Delta P1 Delta 2A Delta 2B的RNA不能复制并形成子代病毒。这些结果表明，2A（pro）是不需要的复制P1无效PV，虽然它是重要的PV RNA复制和诱导CPE。为了知道2A（pro）缺陷型PV是否可以用作载体，检查了含有增强型绿色荧光蛋白（EGFP）编码序列的P1无效PV，该序列具有或不具有2A（pro）。表达荧光蛋白。这一结果表明2A（pro）缺陷型PV可以表达外源基因。
Poliovirus (PV) 2A(pro) has been considered important for PV replication and is known to be toxic to host cells. A 2A(pro)-deficient PV would potentially be less toxic and ideal as a vector. To examine whether 2A(pro) is needed to form progeny virus, a full-length cDNA of dicistronic (dc) PV with (pOME) or without (pOME Delta 2A) 2A(pro) was constructed in the strain PV1(M)OM. RNAs of both pOME and pOME Delta 2A were capable of forming progeny viruses, called OME and OME Delta 2A, respectively. In their ability to induce a cytopathic effect (CPE), the strains ranked as OME Delta 2A < OME is approximately equal to PV1(M) OM. These results suggest that 2A(pro) is not essential for full-length dc PV to form progeny virus and that it contributes to the efficient viral replication and/or induction of a CPE. To clarify whether 2A(pro) is essential for P1-null (lacking the entire coding sequence for capsid proteins) PV, the RNA replication activity of P1-null PV (pOM Delta P1) or P1-null PV without 2A(pro) (pOM Delta P1 Delta 2A) or without both 2A(pro) and 2B (pOM Delta P1 Delta 2A Delta 2B) was examined. The RNAs of pOM Delta P1 and pOM Delta P1 Delta 2A could replicate and form progeny viruses under a trans supply of P1 protein, whereas the RNA of pOM Delta P1 Delta 2A Delta 2B could not. These results suggest that 2A(pro) is not needed for the replication of P1-null PV, although it is important for PV RNA replication and inducing a CPE. To know whether a 2A(pro)-deficient PV can be used as a vector, a P1-null PV containing the enhanced green fluorescent protein (EGFP) coding sequence with or without 2A(pro) was examined. It expressed fluorescent protein. This result suggests that 2A(pro)-deficient PV can express foreign genes.