Roy C, Lebleu B
Laboratoire de Biochimie des Protéines, URA CNRS 1191, Université Montpellier II, France.
Nucleic Acids Res. 1991 Feb 11;19(3):517-24. doi: 10.1093/nar/19.3.517.
The regions of several genes (IFI-56K, HLA-A3, HLA-DR and 6-16) containing the (putative) ISRE (Interferon Stimulatable Response Element) were tested for their ability to be recognized by HeLa cells nuclear extract proteins. In a band shift assay, all probes yielded two B1 and B2 DNA-protein complexes of similar mobilities. Unexpectedly the titration of the B1 complex with a synthetic ISRE core (OL1), promoted the formation of B2. Both the probe and OL1 were recovered in B2. For each probe, the possibility of the part of the sequence involved in B1 complex to form a H-DNA structure with the part of the sequence involved in B2 exists. Such a structure was favored by the colinearity of the pairing regions and requires ATP. Although probes seemed to have a secondary structure, the formal existence of a H-DNA structure has not been demonstrated. Such a model could be extended to other interferon inducible gene promoters and may account for their binding properties and differential inducibility after 5' deletion or point mutations.
对几个含有(假定的)干扰素刺激反应元件(ISRE)的基因(IFI-56K、HLA-A3、HLA-DR和6-16)区域进行了测试,以检测其被HeLa细胞核提取物蛋白识别的能力。在凝胶迁移实验中,所有探针均产生了两个迁移率相似的B1和B2 DNA-蛋白质复合物。出乎意料的是,用合成的ISRE核心(OL1)滴定B1复合物会促进B2的形成。探针和OL1都存在于B2中。对于每个探针,参与B1复合物的序列部分与参与B2的序列部分形成H-DNA结构的可能性是存在的。这种结构受配对区域共线性的青睐,并且需要ATP。尽管探针似乎具有二级结构,但尚未证明H-DNA结构的正式存在。这样的模型可以扩展到其他干扰素诱导基因启动子,并可能解释它们的结合特性以及5'端缺失或点突变后的差异诱导性。