THE E3L GENE OF VACCINIA VIRUS ENCODES AN INHIBITOR OF THE INTERFERON-INDUCED, DOUBLE-STRANDED RNA-DEPENDENT PROTEIN-KINASE
THE E3L GENE OF VACCINIA VIRUS ENCODES AN INHIBITOR OF THE INTERFERON-INDUCED, DOUBLE-STRANDED RNA-DEPENDENT PROTEIN-KINASE
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DOI：
10.1073/pnas.89.11.4825
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发表时间：
1992-06-01
影响因子：
11.1
通讯作者：
JACOBS, BL
中科院分区：
文献类型：
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作者：
CHANG, HW;WATSON, JC;JACOBS, BL
牛痘病毒编码的双链RNA结合蛋白p25先前已被认为与干扰素诱导的双链RNA激活蛋白激酶的抑制有关。在这项研究中，我们已经确定了编码p25的牛痘病毒基因（WR株）。p25的胰凝乳蛋白酶片段的氨基酸序列分析揭示了与牛痘病毒（哥本哈根株）E3 L基因的密切匹配。克隆了WR株E3 L基因，并在COS-1细胞和兔网织红细胞裂解液中进行了体外表达。在每种情况下，都产生了能够与聚（rI）、聚（rC）-琼脂糖结合并与p25特异性抗血清反应的M（r）25，000多肽。此外，COS细胞表达E3 L基因产物抑制干扰素处理的细胞提取物中的双链RNA激活的蛋白激酶的激活。通过用抗p25抗血清吸附去除E3 L编码的产物导致激酶抑制活性的丧失。这些结果表明，牛痘病毒E3 L基因编码p25，并且E3 L基因的产物具有激酶抑制活性。E3 L基因产物的推导的氨基酸序列与蛋白质序列数据库的比较揭示了与人干扰素诱导的双链RNA激活的蛋白激酶密切相关的区域。
A vaccinia virus-encoded double-stranded RNA-binding protein, p25, has been previously implicated in inhibition of the interferon-induced, double-stranded RNA-activated protein kinase. In this study, we have identified the vaccinia viral gene (WR strain) that encodes p25. Amino acid sequence analysis of a chymotryptic fragment of p25 revealed a close match to the vaccinia virus (Copenhagen strain) E3L gene. The WR strain E3L gene was cloned and expressed either in COS-1 cells or in rabbit reticulocyte lysates in vitro. A M(r) 25,000 polypeptide that could bind to poly(rI).poly(rC)-agarose and that reacted with p25-specific antiserum was produced in each case. In addition, COS cells expressing E3L gene products inhibited activation of the double-stranded RNA-activated protein kinase in extracts from interferon-treated cells. Removal of E3L-encoded products by adsorption with anti-p25 antiserum resulted in loss of kinase inhibitory activity. These results demonstrate that the vaccinia virus E3L gene encodes p25 and that the products of the E3L gene have kinase inhibitory activity. Comparison of the deduced amino acid sequence of the E3L gene products with the protein sequence data base revealed a region closely related to the human interferon-induced, double-stranded RNA-activated protein kinase.