Possible role of dimerization in human immunodeficiency virus type 1 genome RNA packaging
Possible role of dimerization in human immunodeficiency virus type 1 genome RNA packaging
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DOI：
10.1128/jvi.77.7.4060-4069.2003
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发表时间：
2003-04-01
影响因子：
5.4
通讯作者：
Shioda, T
中科院分区：
文献类型：
--
作者：
Sakuragi, J;Ueda, S;Shioda, T
二聚体起始位点/二聚体连接序列（DIS/DLS）区域在人类免疫缺陷病毒1型（HIV-1）RNA基因组中被认为在病毒生命周期的各个步骤中发挥重要作用。然而，由于存在一个假定的DIS/DLS区位于的突变信号区（E/psi），很难进行DIS/DLS的突变分析，而不影响RNA包装成病毒粒子。最近，我们证明了病毒RNA中DIS/DLS区域的复制导致病毒粒子中部分单体RNA的产生，表明该区域确实介导了RNA-RNA相互作用。我们利用该系统来评估DIS/DLS在HIV-1基因组的5'区域中的精确位置，同时对RNA包装的影响最小。我们发现U 5/L茎环的整个下茎是包装所必需的，而对二聚体形成重要的区域在U 5/L茎环的下茎内只有10个碱基长。R/U 5茎环是RNA包装所必需的，但完全不适合二聚体形成。SL 1下部茎对于二聚化和包装都是重要的，但令人惊讶的是，环顶部的回文序列的缺失仅部分影响二聚化。这些结果清楚地表明，HIV-1的E/psi比DIS/DLS大得多，并且主要DIS/DLS完全包括在E/psi中。因此，这表明RNA二聚化是RNA包装的一部分，这需要多个步骤。
The dimer initiation site/dimer linkage sequence (DIS/DLS) region in the human immunodeficiency virus type 1 (HIV-1) RNA genome is suggested to play important roles in various steps of the virus life cycle. However, due to the presence of a putative DIS/DLS region located within the encapsidation signal region (E/psi), it is difficult to perform a mutational analysis of DIS/DLS without affecting the packaging of RNA into virions. Recently, we demonstrated that duplication of the DIS/DLS region in viral RNA caused the production of partially monomeric RNAs in virions, indicating that the region indeed mediated RNA-RNA interaction. We utilized this system to assess the precise location of DIS/DLS in the 5' region of the HIV-1 genome with minimum effect on RNA packaging. We found that the entire lower stem of the U5/L stem-loop was required for packaging, whereas the region important for dimer formation was only 10 bases long Within the lower stem of the U5/L stem-loop. The R/U5 stem-loop was required for RNA packaging but was completely dispensable for dimer formation. The SL1 lower stem was important for both dimerization and packaging, but surprisingly, deletion of the palindromic sequence at the top of the loop only partially affected dimerization. These results clearly indicated that the E/psi of HIV-1 is much larger than the DIS/DLS and that the primary DIS/DLS is completely included in the E/psi. Therefore, it is suggested that RNA dimerization is a part of RNA packaging, which requires multiple steps.