Raghunathan Devanathan, Sánchez-Pedregal Víctor M, Junker Jochen, Schwiegk Claudia, Kalesse Markus, Kirschning Andreas, Carlomagno Teresa
Department of NMR-based Structural Biology, The Max Planck Institute for Biophysical Chemistry, Am Fassberg, 11 D-37077 Göttingen, Germany.
Nucleic Acids Res. 2006 Jul 19;34(12):3599-608. doi: 10.1093/nar/gkl494. Print 2006.
The formation of the Tat-protein/TAR-RNA complex is a crucial step in the regulation of human immunodeficiency virus (HIV)-gene expression. To obtain full-length viral transcripts the Tat/TAR complex has to recruit the positive transcription elongation factor complex (P-EFTb), which interacts with TAR through its cyclin T1 (CycT1) component. Mutational studies identified the TAR hexanucleotide loop as a crucial region for contacting CycT1. Interfering with the interaction between the Tat/CycT1 complex and the TAR-RNA is an attractive strategy for the design of anti-HIV drugs. Positively charged molecules, like aminoglycosides or peptidomimetics, bind the TAR-RNA, disrupting the Tat/TAR complex. Here, we investigate the complex between the HIV-2 TAR-RNA and a neooligoaminodeoxysaccharide by NMR spectroscopy. In contrast to other aminoglycosides, this novel aminoglycoside analogue contacts simultaneously the bulge residues required for Tat binding and the A35 residue of the hexanucleotide loop. Upon complex formation, the loop region undergoes profound conformational changes. The novel binding mode, together with the easy accessibility of derivatives for the neooligoaminodeoxysaccharide, could open the way to the design of a new class of TAR-RNA binders, which simultaneously inhibit the formation of both the Tat/TAR binary complex and the Tat/TAR/CycT1 ternary complex by obstructing both the bulge and loop regions of the RNA.
Tat 蛋白/TAR-RNA 复合物的形成是人类免疫缺陷病毒(HIV)基因表达调控中的关键步骤。为了获得全长病毒转录本,Tat/TAR 复合物必须招募正性转录延伸因子复合物(P-EFTb),该复合物通过其细胞周期蛋白 T1(CycT1)组分与 TAR 相互作用。突变研究确定 TAR 六核苷酸环是与 CycT1 接触的关键区域。干扰 Tat/CycT1 复合物与 TAR-RNA 之间的相互作用是设计抗 HIV 药物的一种有吸引力的策略。带正电荷的分子,如氨基糖苷类或肽模拟物,可结合 TAR-RNA,破坏 Tat/TAR 复合物。在此,我们通过核磁共振光谱研究了 HIV-2 TAR-RNA 与一种新的低聚氨基脱氧糖之间的复合物。与其他氨基糖苷类不同,这种新型氨基糖苷类似物同时接触 Tat 结合所需的凸起残基和六核苷酸环的 A35 残基。复合物形成后,环区域会发生深刻的构象变化。这种新型结合模式,以及新的低聚氨基脱氧糖衍生物易于获得的特点,可能为设计一类新的 TAR-RNA 结合剂开辟道路,这类结合剂通过阻碍 RNA 的凸起和环区域,同时抑制 Tat/TAR 二元复合物和 Tat/TAR/CycT1 三元复合物的形成。