Division of Structural Biology, Wellcome Trust Centre for Human Genetics, Oxford University, Roosevelt Drive, Oxford OX3 7BN, UK.
Proc Natl Acad Sci U S A. 2010 Mar 30;107(13):5810-4. doi: 10.1073/pnas.0914946107. Epub 2010 Mar 15.
HIV-1 Rev is a small regulatory protein that mediates the nuclear export of viral mRNAs, an essential step in the HIV replication cycle. In this process Rev oligomerizes in association with a highly structured RNA motif, the Rev response element. Crystallographic studies of Rev have been hampered by the protein's tendency to aggregate, but Rev has now been found to form a stable soluble equimolar complex with a specifically engineered monoclonal Fab fragment. We have determined the structure of this complex at 3.2 A resolution. It reveals a molecular dimer of Rev, bound on either side by a Fab, where the ordered portion of each Rev monomer (residues 9-65) contains two coplanar alpha-helices arranged in hairpin fashion. Subunits dimerize through overlapping of the hairpin prongs. Mating of hydrophobic patches on the outer surface of the dimer is likely to promote higher order interactions, suggesting a model for Rev oligomerization onto the viral RNA.
HIV-1 Rev 是一种小的调节蛋白,介导病毒 mRNA 的核输出,这是 HIV 复制周期中的一个关键步骤。在这个过程中,Rev 与高度结构化的 RNA 基序(Rev 反应元件)结合形成寡聚体。Rev 的晶体学研究受到蛋白质聚集倾向的阻碍,但现在已经发现 Rev 与专门设计的单克隆 Fab 片段形成稳定的可溶性等摩尔复合物。我们已经确定了该复合物在 3.2Å分辨率下的结构。它揭示了 Rev 的分子二聚体,两侧由 Fab 结合,每个 Rev 单体的有序部分(残基 9-65)包含两个共面的α-螺旋以发夹形式排列。亚基通过发夹叉的重叠二聚化。二聚体外部表面上的疏水斑的配对可能促进更高阶的相互作用,这表明 Rev 与病毒 RNA 形成寡聚体的模型。