Binding properties of human immunodeficiency virus type‐2 (HIV‐2) rna corresponding to the packaging signal to its nucleocapsid protein
Binding properties of human immunodeficiency virus type‐2 (HIV‐2) rna corresponding to the packaging signal to its nucleocapsid protein
复制标题
人类免疫缺陷病毒 2 型 (HIV-2) rna 的结合特性与其核衣壳蛋白的包装信号相对应
DOI：
10.1080/15216549600201502
复制
发表时间：
1996
期刊：
影响因子：
4.6
通讯作者：
H. Tozawa
中科院分区：
文献类型：
--
作者：
Tomonori Tsukahara;H. Komatsu;M. Kubo;F. Obata;H. Tozawa
人类免疫缺陷病毒1型（HIV‐1）的核衣壳蛋白（NC）含有两个锌指基序（ZFMs），并特异性结合位于病毒基因组RNA 5 '先导序列中位于第一剪接供体和gag启动密码子（AUG）之间的包装信号。在这项研究中，我们使用体外合成的32P标记和未标记的rna进行竞争紫外线（UV）交联实验，分析了HIV‐2相应区域（3区）与其NC蛋白（NCp8）结合的特异性，这些rna对应于引物结合位点和gag AUG（1区）之间的序列。添加未标记的1区和3区RNA可特异性抑制32P标记的1区RNA与NCp8的结合，而使用NCp8的缺失突变肽则未检测到特异性结合。这些发现表明，与HIV‐2 NCp8特异性结合的区域位于第一个剪接供体和5 '先导序列中的gag AUG之间，NCp8是负责与HIV‐2 RNA第一个剪接供体位点下游区域特异性结合的最小结合区域。
The nucleocapsid (NC) protein of human immunodeficiency virus type‐1 (HIV‐1) contains two zinc finger motifs (ZFMs), and binds specifically to the packaging signal which is located in the 5′ leader sequence of the viral genomic RNA between the first splice donor and the gag initiator codon (AUG). In this study, we analyzed the specificity of the binding of the corresponding region of HIV‐2 (Region 3) to its NC protein (NCp8), by performing a competitive ultraviolet (UV) cross‐linking assay using in vitro‐synthesized 32P‐labeled and unlabeled RNAs corresponding to a sequence between the primer binding site and the gag AUG (Region 1). Binding of 32P‐labeled Region 1 RNA to NCp8 was inhibited specifically by adding unlabeled Region 1 and 3 RNAs and no specific binding was detected using deletion mutant peptides of NCp8. These findings suggest that the region(s) which bind(s) specifically to HIV‐2 NCp8 lie(s) between the first splice donor and the gag AUG in the 5′ leader sequence and that NCp8 is the minimum binding region responsible for the specific binding of the region downstream of the first splice donor site of HIV‐2 RNA.