Schaal Thomas D, Mallet William G, McMinn Dustin L, Nguyen Nam V, Sopko Michelle M, John Sam, Parekh Bhavin S
Laboratory of Receptor Biology and Gene Expression, Center for Cancer Research, Building 41, Room B307, NCI, NIH, Bethesda, MD 20892-5055, USA.
Nucleic Acids Res. 2003 Feb 15;31(4):1282-91. doi: 10.1093/nar/gkg206.
Polyamides are a class of heterocyclic small molecules with the potential of controlling gene expression by binding to the minor groove of DNA in a sequence-specific manner. To evaluate the feasibility of this class of compounds as antiviral therapeutics, molecules were designed to essential sequence elements occurring numerous times in the HPV genome. This sequence element is bound by a virus-encoded transcription and replication factor E2, which binds to a 12 bp recognition site as a homodimeric protein. Here, we take advantage of polyamide:DNA and E2:DNA co-crystal structural information and advances in polyamide synthetic chemistry to design tandem hairpin polyamides that are capable of displacing the major groove-binding E2 homodimer from its DNA binding site. The binding of tandem hairpin polyamides and the E2 DNA binding protein to the DNA site is mutually exclusive even though the two ligands occupy opposite faces of the DNA double helix. We show with circular permutation studies that the tandem hairpin polyamide prevents the intrinsic bending of the E2 DNA site important for binding of the protein. Taken together, these results illustrate the feasibility of inhibiting the binding of homodimeric, major groove-binding transcription factors by altering the local DNA geometry using minor groove-binding tandem hairpin polyamides.
聚酰胺是一类杂环小分子,具有通过以序列特异性方式与DNA小沟结合来控制基因表达的潜力。为了评估这类化合物作为抗病毒治疗药物的可行性,设计了针对人乳头瘤病毒(HPV)基因组中多次出现的必需序列元件的分子。该序列元件由病毒编码的转录和复制因子E2结合,E2作为同二聚体蛋白与一个12bp的识别位点结合。在此,我们利用聚酰胺:DNA和E2:DNA共晶体结构信息以及聚酰胺合成化学的进展,设计出能够将结合于大沟的E2同二聚体从其DNA结合位点置换出来的串联发夹聚酰胺。尽管两种配体占据DNA双螺旋的相对面,但串联发夹聚酰胺与E2 DNA结合蛋白对DNA位点的结合是相互排斥的。我们通过环形置换研究表明,串联发夹聚酰胺可阻止对蛋白结合很重要的E2 DNA位点的固有弯曲。综上所述,这些结果说明了使用结合小沟的串联发夹聚酰胺改变局部DNA几何形状来抑制同二聚体、结合大沟的转录因子结合的可行性。