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人双链RNA激活的p68激酶在鼠细胞中的组成型表达介导真核起始因子2的磷酸化并对脑心肌炎病毒生长产生部分抗性。

Constitutive expression of human double-stranded RNA-activated p68 kinase in murine cells mediates phosphorylation of eukaryotic initiation factor 2 and partial resistance to encephalomyocarditis virus growth.

作者信息

Meurs E F, Watanabe Y, Kadereit S, Barber G N, Katze M G, Chong K, Williams B R, Hovanessian A G

机构信息

Unit of Virology and of Cellular Immunology (UA CNRS 1157), Institut Pasteur, Paris, France.

出版信息

J Virol. 1992 Oct;66(10):5805-14. doi: 10.1128/JVI.66.10.5805-5814.1992.

DOI:10.1128/JVI.66.10.5805-5814.1992
PMID:1382142
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC241456/
Abstract

The cDNA encoding interferon-induced human double-stranded RNA-activated p68 kinase was expressed in murine NIH 3T3 cells by using the pcDNA1/neo vector. Several stable clones were selected which expressed either the wild-type kinase or an inactive mutant possessing a single amino acid substitution in the invariant lysine 296 in the catalytic domain II. The transfected wild-type kinase showed properties similar to those of the natural kinase, such as subcellular ribosomal localization and dependence on double-stranded RNA for autophosphorylation. Upon infection with encephalomyocarditis virus (EMCV), wild-type- but not mutant-expressing clones were found to partially resist virus growth. Such natural antiviral activity was virus specific, since no inhibition was observed in the case of vesicular stomatitis virus infection. In accord with EMCV inhibition, the wild-type p68 kinase was found to be highly phosphorylated during infection. Furthermore, its natural substrate, the small subunit of protein synthesis initiation factor eIF2, was phosphorylated. These results demonstrate that p68 kinase is activated during EMCV infection, leading to reduced virus production.

摘要

利用pcDNA1/neo载体，将编码干扰素诱导的人双链RNA激活的p68激酶的cDNA在鼠NIH 3T3细胞中进行表达。挑选出了几个稳定克隆，这些克隆表达野生型激酶或在催化结构域II的不变赖氨酸296处具有单个氨基酸取代的无活性突变体。转染的野生型激酶表现出与天然激酶相似的特性，如亚细胞核糖体定位以及自磷酸化对双链RNA的依赖性。在用脑心肌炎病毒（EMCV）感染后，发现表达野生型而非突变体的克隆能部分抵抗病毒生长。这种天然抗病毒活性具有病毒特异性，因为在水疱性口炎病毒感染的情况下未观察到抑制作用。与EMCV抑制作用一致，发现野生型p68激酶在感染期间高度磷酸化。此外，其天然底物——蛋白质合成起始因子eIF2的小亚基也被磷酸化。这些结果表明，p68激酶在EMCV感染期间被激活，从而导致病毒产生减少。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/29296df245b6/jvirol00041-0117-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/8ebe3db022b8/jvirol00041-0113-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/deb1feb160c0/jvirol00041-0114-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/35475824bfe7/jvirol00041-0116-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/29296df245b6/jvirol00041-0117-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/8ebe3db022b8/jvirol00041-0113-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/deb1feb160c0/jvirol00041-0114-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/35475824bfe7/jvirol00041-0116-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/828d/241456/29296df245b6/jvirol00041-0117-a.jpg

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