RECOGNITION OF THE HIGH-AFFINITY BINDING-SITE IN REV-RESPONSE ELEMENT RNA BY THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REV PROTEIN
RECOGNITION OF THE HIGH-AFFINITY BINDING-SITE IN REV-RESPONSE ELEMENT RNA BY THE HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 REV PROTEIN
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DOI：
10.1093/nar/20.24.6465
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发表时间：
1992-12-25
影响因子：
14.9
通讯作者：
GAIT, MJ
中科院分区：
文献类型：
--
作者：
IWAI, S;PRITCHARD, C;GAIT, MJ
人类免疫缺陷病毒1型rev蛋白以高亲和力结合在茎环II的rev-response元件（RRE） RNA中的泡结构上。在最初的相互作用之后，额外的rev分子以有序的组装过程与RRE RNA结合，这需要一个功能性的泡结构，因为泡序列中的突变会降低rev亲和力，从而阻止多个复合物的形成。我们利用合成化学方法来表征rev蛋白与其高亲和力结合位点之间的相互作用。一个最小的合成双工RNA （RBC6）携带气泡和12个侧翼碱基对，能够以1对1的化学计量和高亲和力结合rev。当气泡结构插入到携带较长双链RNA的合成RNA分子中时，rev会形成额外的复合物，类似于在RRE RNA中观察到的多聚体。我们还研究了rev与RBC6类似物结合的能力，这些类似物在核苷残基的碱基和糖基上含有官能团修饰。结果提供了强有力的证据，表明气泡结构包含非沃森-克里克G:G和G:A碱基对的特定构型，并表明rev对RRE RNA的高亲和力识别需要与扭曲RNA结构主槽中的功能基团形成氢键。
The Human Immunodeficiency Virus type-1 rev protein binds with high affinity to a bubble structure located within the rev-response element (RRE) RNA in stem-loop II. After this initial interaction, additional rev molecules bind to the RRE RNA in an ordered assembly process which requires a functional bubble structure, since mutations in the bubble sequence that reduce rev affinity block multiple complex formation. We have used synthetic chemistry to characterize the interaction between rev protein and its high affinity binding site. A minimal synthetic duplex RNA (RBC6) carrying the bubble and 12 flanking base pairs is able to bind rev with 1 to 1 stoichiometry and with high affinity. When the bubble structure is inserted into synthetic RNA molecules carrying longer stretches of flanking double-stranded RNA, rev forms additional complexes resembling the multimers observed with the RRE RNA. The ability of rev to bind to RBC6 analogues containing functional group modifications on base and sugar moieties of nucleoside residues was also examined. The results provide strong evidence that the bubble structure contains specific configurations of non-Watson-Crick G:G and G:A base pairs and suggest that high affinity recognition of RRE RNA by rev requires hydrogen bonding to functional groups in the major groove of a distorted RNA structure.