Replication of human immunodeficiency viruses engineered with heterologous tat-transactivation response element interactions
Replication of human immunodeficiency viruses engineered with heterologous tat-transactivation response element interactions
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DOI：
10.1128/jvi.77.3.1984-1991.2003
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发表时间：
2003-02-01
影响因子：
5.4
通讯作者：
Frankel, AD
中科院分区：
文献类型：
--
作者：
Xie, BD;Wainberg, MA;Frankel, AD
人类免疫缺陷病毒（HIV）和相关的牛慢病毒牛免疫缺陷病毒（BIV）和杰布拉纳病病毒（JDV）利用病毒达特蛋白来激活病毒转录。达特蛋白的富含精氨酸的RNA结合结构域与位于病毒mRNA 5'端的同源反式激活反应元件（TAR）RNA发夹结合，导致RNA聚合酶II的持续合成能力增强。先前已经表明，I型HIV（HIV-1）达特需要细胞周期蛋白T1蛋白用于高亲和力RNA结合，而BIV达特和JDV达特自身以高亲和力结合，并且当与RNA复合时采用不同的β-发夹构象。在这里，我们已经将BIV和JDV Tat-TAR相互作用工程化到HIV-1中，并表明异源相互作用支持病毒复制，与它们的RNA结合亲和力很好地相关。用能够结合所有三种达特蛋白的变体TAR工程化的病毒与任何蛋白有效复制。在一种含有既不相互作用也不有效复制的非同源Tat-TAR对（HIV-1 TAR和BIV达特）的病毒中，分离出TAR突变产生功能性BIV达特结合位点的病毒回复突变体。我们的研究结果支持这样的观点，即TAR结构的增量变化可以为发展新的Tat-TAR复合物提供途径，同时保持活跃的病毒复制。
Human immunodeficiency viruses (HIVs) and the related bovine lentiviruses bovine immunodeficiency virus (BIV) and Jembrana disease virus (JDV) utilize the viral Tat protein to activate viral transcription. The arginine-rich RNA-binding domains of the Tat proteins bind to their cognate transactivation response element (TAR) RNA hairpins located at the 5' ends of the viral mRNAs, resulting in enhanced processivity of RNA polymerase II. It has previously been shown that HIV type I (HIV-1) Tat requires the cellular cyclin T1 protein for high-affinity RNA binding whereas BIV Tat and JDV Tat bind with high affinity on their own and adopt distinct beta-hairpin conformations when complexed to RNA. Here we have engineered the BIV and JDV Tat-TAR interactions into HIV-1 and show that the heterologous interactions support viral replication, correlating well with their RNA-binding affinities. Viruses engineered with a variant TAR able to bind all three Tat proteins replicate efficiently with any of the proteins. In one virus containing a noncognate Tat-TAR pair that neither interacts nor efficiently replicates (HIV-1 TAR and BIV Tat), viral revertants were isolated in which TAR had become mutated to generate a functional BIV Tat binding site. Our results support the view that incremental changes to TAR structure can provide routes for evolving new Tat-TAR complexes while maintaining active viral replication.