Lochrie M A, Waugh S, Pratt D G, Clever J, Parslow T G, Polisky B
NeXstar Pharmaceuticals, Inc., 2860 Wilderness Place, Boulder, CO 80301, USA.
Nucleic Acids Res. 1997 Jul 15;25(14):2902-10. doi: 10.1093/nar/25.14.2902.
RNA ligands that bind to the human immunodeficiency virus type-1 (HIV-1) gag polyprotein with 10(-9) M affinity were isolated from a complex pool of RNAs using an in vitro selection method. The ligands bind to two different regions within gag, either to the matrix protein or to the nucleocapsid protein. Binding of a matrix ligand to gag did not interfere with the binding of a nucleocapsid ligand, and binding of a nucleocapsid ligand to gag did not interfere with the binding of a matrix ligand. However, binding of a nucleocapsid ligand to gag did interfere with binding of an RNA containing the HIV-1 RNA packaging element (psi), even though the sequence of the nucleocapsid ligand is not similar topsi. The minimal sequences required for the ligands to bind to matrix or nucleocapsid were determined. Minimal nucleocapsid ligands are predicted to form a stem-loop structure that has a self-complementary sequence at one end. Minimal matrix ligands are predicted to form a different stem-loop structure that has a CAARU loop sequence. The properties of these RNA ligands may provide tools for studying RNA interactions with matrix and nucleocapsid, and a novel method for inhibiting HIV replication.
利用体外筛选方法,从复杂的RNA文库中分离出了与人类免疫缺陷病毒1型(HIV-1)gag多聚蛋白以10⁻⁹ M亲和力结合的RNA配体。这些配体与gag内的两个不同区域结合,要么与基质蛋白结合,要么与核衣壳蛋白结合。基质配体与gag的结合不干扰核衣壳配体与gag的结合,核衣壳配体与gag的结合也不干扰基质配体与gag的结合。然而,核衣壳配体与gag的结合确实会干扰含有HIV-1 RNA包装元件(ψ)的RNA的结合,尽管核衣壳配体的序列与ψ不相似。确定了配体与基质或核衣壳结合所需的最小序列。预测最小的核衣壳配体形成一种茎环结构,其一端具有自我互补序列。预测最小的基质配体形成一种不同的茎环结构,其具有CAARU环序列。这些RNA配体的特性可能为研究RNA与基质和核衣壳的相互作用提供工具,并为抑制HIV复制提供一种新方法。