Suzow J, Friedman A D
Division of Pediatric Oncology, Johns Hopkins Oncology Center, Johns Hopkins University, Baltimore, Maryland 21287.
Mol Cell Biol. 1993 Apr;13(4):2141-51. doi: 10.1128/mcb.13.4.2141-2151.1993.
The myeloperoxidase (MPO) gene is expressed specifically in myeloid cells. There is significant homology between the murine and human MPO genes in the 1.6-kb region located upstream of the murine MPO transcription initiation sites. 5',3', and internal deletions of this DNA segment localized several cis-acting DNA elements in the murine MPO promoter which are functional in 32D cl3 cells, a murine myeloblast cell line which expresses MPO. These DNA elements did not function well in mouse L-cell fibroblasts. Additional mutagenesis of the most active promoter region allowed the delimitation of a functional 20-bp segment. Mutation of the enhancer core motif within this segment was functionally deleterious, and an oligonucleotide containing these base pairs increased the activity of a minimal promoter. This same oligonucleotide, but not a mutant variant, could bind a set of nuclear proteins, myeloid nuclear factors 1 alpha and 1 beta (MyNF1 alpha and -1 beta), present in 32D cl3 cells but absent from L cells, murine erythroleukemia cells, and SP2 lymphoid cells.
髓过氧化物酶(MPO)基因在髓样细胞中特异性表达。在小鼠MPO转录起始位点上游1.6 kb区域内,小鼠和人类MPO基因之间存在显著同源性。该DNA片段的5'、3'和内部缺失定位了小鼠MPO启动子中的几个顺式作用DNA元件,这些元件在32D cl3细胞(一种表达MPO的小鼠成髓细胞系)中具有功能。这些DNA元件在小鼠L细胞成纤维细胞中功能不佳。对活性最高的启动子区域进行进一步诱变,确定了一个功能性的20 bp片段。该片段内增强子核心基序的突变在功能上是有害的,并且含有这些碱基对的寡核苷酸增加了最小启动子的活性。这个相同的寡核苷酸,但不是突变变体,可以结合一组核蛋白,即存在于32D cl3细胞中但不存在于L细胞、小鼠红白血病细胞和SP2淋巴细胞中的髓核因子1α和1β(MyNF1α和-1β)。