Department of Molecular, Cellular and Biomedical Studies, City University of New York School of Medicine and B.S.-M.D. Program, Harris Hall, 160 Convent Avenue, New York, NY 10031, USA.
City University of New York Graduate Center, Programs in Biochemistry and Biology, 365 Fifth Ave, New York, NY 10016, USA.
Biomolecules. 2020 Mar 7;10(3):417. doi: 10.3390/biom10030417.
does not contain a p53 homolog. Utilizing this yeast as an in vivo test tube model, our aim was to investigate if a yeast protein would show p53 DNA binding homology. Electrophoretic mobility shift analyses revealed the formation of specific DNA-protein complexes consisting of nuclear protein(s) and oligonucleotides containing p53 DNA binding sites. A p53 binding site factor (Scp53BSF) bound to a p53 synthetic DNA-consensus sequence (SCS) and a p53 binding-site sequence from the oncogene. The complexes were of comparable size. Like mammalian p53, the affinity of Scp53BSF for the SCS oligonucleotide was higher than for the MDM2 oligonucleotide. Binding of Scp53BSF to the SCS and MDM2 oligonucleotides was strongly competed by unlabeled oligonucleotides containing mammalian p53 sites, but very little by a mutated site oligonucleotide. Importantly, Scp53BSF-DNA binding activity was significantly induced in extracts from cells with DNA damage. This resulted in dose-dependent coordinated activation of transcription when using p53-binding site reporter constructs. An ancient p53-like DNA binding protein may have been found, and activation of DNA-associated factors to p53 response elements may have functions not yet determined.
它不含有 p53 同源物。我们利用这种酵母作为体内试管模型，旨在研究酵母蛋白是否会显示出与 p53 DNA 结合的同源性。电泳迁移率变动分析显示，由核蛋白和含有 p53 DNA 结合位点的寡核苷酸组成的特定 DNA-蛋白复合物的形成。p53 结合位点因子（Scp53BSF）与 p53 合成 DNA-共有序列（SCS）和来自癌基因的 p53 结合位点序列结合。这些复合物的大小相当。与哺乳动物 p53 一样，Scp53BSF 对 SCS 寡核苷酸的亲和力高于对 MDM2 寡核苷酸的亲和力。Scp53BSF 与 SCS 和 MDM2 寡核苷酸的结合可被含有哺乳动物 p53 位点的未标记寡核苷酸强烈竞争，但被突变位点寡核苷酸的竞争很小。重要的是，来自具有 DNA 损伤的细胞的提取物中 Scp53BSF-DNA 结合活性显著诱导。当使用 p53 结合位点报告基因构建体时，这导致了剂量依赖性的转录协调激活。可能已经发现了一种古老的 p53 样 DNA 结合蛋白，并且 DNA 相关因子对 p53 反应元件的激活可能具有尚未确定的功能。
