Pietrowski D, Durante M J, Liebstein A, Schmitt-John T, Werner T, Graw J
GSF-Forschungszentrum für Umwelt und Gesundheit, Institut für Säugetiergenetik, Neuherberg, Germany.
Gene. 1994 Jul 8;144(2):171-8. doi: 10.1016/0378-1119(94)90375-1.
The promoter of the murine gamma E-crystallin (gamma E-Cry) encoding gene (gamma E-cry) was analyzed for specific interactions with lenticular proteins in a gel-retardation assay. A 21-bp fragment immediately downstream of the transcription initiation site (DOTIS) is demonstrated to be responsible for specific interactions with lens extracts. The DOTIS-binding protein(s) accept only the sense DNA strand as target; anti-sense or double-stranded DNA do not interact with these proteins. The DOTIS sequence element is highly conserved among the murine gamma D-, gamma E- and gamma F-cry and is present at comparable positions in the orthologous rat genes. Only a weak or even no protein-binding activity is observed if a few particular bases are changed, as in the rat gamma A-, gamma C- and gamma E-cry elements. DOTIS-binding proteins were found in commercially available bovine alpha-Cry preparations. The essential participation of alpha-Cry in the DNA-binding protein complex was confirmed using alpha-Cry-specific monoclonal antibody. The results reported here point to a novel function of alpha-Cry besides the structural properties in the lens.
通过凝胶阻滞试验分析了小鼠γE-晶状体蛋白（γE-Cry）编码基因（γE-cry）的启动子与晶状体蛋白的特异性相互作用。转录起始位点下游紧邻的一个21 bp片段（DOTIS）被证明负责与晶状体提取物的特异性相互作用。DOTIS结合蛋白仅将有义DNA链作为靶标；反义或双链DNA不与这些蛋白相互作用。DOTIS序列元件在小鼠γD-、γE-和γF-晶状体蛋白中高度保守，并且在大鼠直系同源基因的相应位置也存在。如果少数特定碱基发生改变，如在大鼠γA-、γC-和γE-晶状体蛋白元件中，仅观察到弱的甚至没有蛋白结合活性。在市售的牛α-晶状体蛋白制剂中发现了DOTIS结合蛋白。使用α-晶状体蛋白特异性单克隆抗体证实了α-晶状体蛋白在DNA结合蛋白复合物中的重要参与。此处报道的结果表明α-晶状体蛋白在晶状体中除了具有结构特性外还具有新功能。
