Regulation of human immunodeficiency virus replication by 2′,5′-oligoadenylate-dependent RNase L
Regulation of human immunodeficiency virus replication by 2′,5′-oligoadenylate-dependent RNase L
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DOI：
10.1128/jvi.72.2.1146-1152.1998
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发表时间：
1998-02-01
影响因子：
5.4
通讯作者：
Silverman, RH
中科院分区：
文献类型：
--
作者：
Maitra, RK;Silverman, RH
2‘5’连接的寡腺苷(2 5A)激活核糖核酸酶(RNase)是干扰素抗病毒作用的途径之一。为了确定2-5A系统在人类免疫缺陷病毒1型复制控制中的作用，用人核糖核酸酶L基因替换了人类免疫缺陷病毒1nef基因的一段。含正义定向核糖核酸酶L基因的HIV-1前病毒在Jurkat细胞中可诱导核糖核酸酶L的表达增加，病毒复制受到500~1000倍的抑制，持续约2周。随后，随着病毒产量的增加，核糖核酸酶L基因的部分缺失发生了。核糖核酸酶L的抗HIV活性与HIV-1RNA的减少有关，并伴随着DNA片段化加速细胞死亡。编码核糖核酸酶L的HIV-1在外周血淋巴细胞中的复制也受到一过性抑制。相反，含有反向核糖核酸酶L基因的重组HIV导致核糖核酸酶L水平下降，HIV产量增加，并降低了α干扰素在PBL和Jurkat细胞中的抗HIV作用。为了获得核糖核酸酶L的组成性和持续性表达，将HIV-1前病毒DNA与含有驱动核糖核酸酶L基因表达的巨细胞病毒启动子的载体共转染Jurkat细胞。核糖核酸酶L质粒抑制HIV-1复制八倍，而反义核糖核酸酶L构建的病毒产量增加两倍。这些发现表明，核糖核酸酶L可以严重损害艾滋病毒的复制，并提示2-5A系统参与了α干扰素的抗艾滋病毒作用。
Activation of RNase by 2'5'-linked oligoadenylates (2-5A) is one of the antiviral pathways in interferon action. To determine the involvement of the 2-5A system in the control of human immunodeficiency virus type 1 (HIV-1) replication, a segment of the HIV-1 nef gene was replaced with human RNase L cDNA. HIV-1 provirus containing sense orientation RNase L cDNA caused increased expression of RNase L and 500- to 1,000-fold inhibition of virus replication in Jurkat cells for a period of about 2 weeks. Subsequently, a partial deletion of the RNase L cDNA which coincided with increases in virus production occurred. The anti-HIV activity of RNase L correlated with decreases in HIV-1 RNA with an acceleration in cell death accompanied by DNA fragmentation. Replication of HIV-1 encoding RNase L was also transiently suppressed in peripheral blood lymphocytes (PBL). In contrast, recombinant HIV containing reverse orientation RNase L cDNA caused decreased levels of RNase L, increases in HIV yields, and reductions in the anti-HIV effect of alpha interferon in PBL and in Jurkat cells. To obtain constitutive and continuous expression of RNase L cDNA, Jurkat cells were cotransfected with HIV-1 proviral DNA and with plasmid containing a cytomegalovirus promoter driving expression of RNase L cDNA. The RNase L plasmid suppressed HIV-1 replication by eightfold, while an antisense RNase L construct enhanced virus production by twofold. These findings demonstrate that RNase L can severely impair HIV replication and suggest involvement of the 2-5A system in the anti-HIV effect of alpha interferon.