Rauscher F J, Morris J F, Tournay O E, Cook D M, Curran T
Wistar Institute, Philadelphia, PA 19104.
Science. 1990 Nov 30;250(4985):1259-62. doi: 10.1126/science.2244209.
The Wilms' tumor locus (WTL) at 11p13 contains a gene that encodes a zinc finger-containing protein that has characteristics of a DNA-binding protein. However, binding of this protein to DNA in a sequence-specific manner has not been demonstrated. A synthetic gene was constructed that contained the zinc finger region, and the protein was expressed in Escherichia coli. The recombinant protein was used to identify a specific DNA binding site from a pool of degenerate oligonucleotides. The binding sites obtained were similar to the sequence recognized by the early growth response-1 (EGR-1) gene product, a zinc finger-containing protein that is induced by mitogenic stimuli. A mutation in the zinc finger region of the protein originally identified in a Wilms' tumor patient abolished its DNA-binding activity. These results suggest that the WTL protein may act at the DNA binding site of a growth factor-inducible gene and that loss of DNA-binding activity contributes to the tumorigenic process.
位于11p13的威尔姆斯瘤基因座（WTL）包含一个编码含锌指蛋白的基因，该蛋白具有DNA结合蛋白的特征。然而，尚未证实该蛋白以序列特异性方式与DNA结合。构建了一个包含锌指区域的合成基因，并在大肠杆菌中表达该蛋白。重组蛋白用于从一组简并寡核苷酸中鉴定特定的DNA结合位点。获得的结合位点与早期生长反应-1（EGR-1）基因产物识别的序列相似，EGR-1是一种含锌指蛋白，由促有丝分裂刺激诱导产生。最初在一名威尔姆斯瘤患者中鉴定出的该蛋白锌指区域的突变消除了其DNA结合活性。这些结果表明，WTL蛋白可能作用于生长因子诱导基因的DNA结合位点，并且DNA结合活性的丧失有助于肿瘤发生过程。
