Brys R, Nelles L, van der Schueren E, Silvestre N, Huylebroeck D, Remacle J E
Laboratory of Molecular Biology (CELGEN), University of Leuven, Belgium.
DNA Cell Biol. 1998 Apr;17(4):349-58. doi: 10.1089/dna.1998.17.349.
We have analyzed the transcriptional activity of the human plasminogen activator inhibitor-1 promoter in the fission yeast Schizosaccharomyces pombe. This promoter is active in S. pombe, and the initiation site of transcription corresponds to the site identified previously in mammalian cells. Mutations in the AP-1-binding site (PAI-1 A box) or the HLTF-binding site (the B box), which reduced the basal and phorbol ester-induced levels of PAI-1 expression in human cells, also decreased the transcriptional activity in S. pombe. Gel retardation assays showed that an S. pombe protein binds specifically to this B box element and displays the same B box sequence requirement as HLTF. Furthermore, this yeast protein binds specifically to other HLTF-binding sites in the human immunodeficiency virus-1 long terminal repeat (LTR) and the simian virus 40 (SV40) enhancer. The B box (but not a mutated B box) strongly stimulated transcription when combined with adh downstream promoter elements, indicating that the S. pombe B box-binding protein, like HLTF, is a transcriptional activator. We conclude that the transcriptional activity of the nonviral PAI-1 promoter is controlled by the same promoter elements in S. pombe as in mammalian cells. In addition, mammalian trans-acting factors that bind to these promoter elements were shown to have counterparts with conserved DNA-binding activity in S. pombe. These results further illustrate the conservation of the mechanism of transcription between mammalian cells and fission yeast.
我们分析了人纤溶酶原激活物抑制剂-1(PAI-1)启动子在裂殖酵母粟酒裂殖酵母中的转录活性。该启动子在粟酒裂殖酵母中具有活性,转录起始位点与先前在哺乳动物细胞中确定的位点相对应。AP-1结合位点(PAI-1 A盒)或HLTF结合位点(B盒)中的突变,这些突变降低了人细胞中PAI-1表达的基础水平和佛波酯诱导水平,也降低了粟酒裂殖酵母中的转录活性。凝胶阻滞分析表明,一种粟酒裂殖酵母蛋白特异性结合该B盒元件,并显示出与HLTF相同的B盒序列要求。此外,这种酵母蛋白特异性结合人免疫缺陷病毒1型长末端重复序列(LTR)和猿猴病毒40(SV40)增强子中的其他HLTF结合位点。当与adh下游启动子元件结合时,B盒(而非突变的B盒)强烈刺激转录,表明粟酒裂殖酵母B盒结合蛋白与HLTF一样,是一种转录激活因子。我们得出结论,非病毒PAI-1启动子的转录活性在粟酒裂殖酵母中与在哺乳动物细胞中受相同的启动子元件控制。此外,与这些启动子元件结合的哺乳动物反式作用因子在粟酒裂殖酵母中具有具有保守DNA结合活性的对应物。这些结果进一步说明了哺乳动物细胞和裂殖酵母之间转录机制的保守性。