Iyer V, Struhl K
Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
EMBO J. 1995 Jun 1;14(11):2570-9. doi: 10.1002/j.1460-2075.1995.tb07255.x.
Many yeast promoters contain homopolymeric dA:dT sequences that affect nucleosome formation in vitro and are required for wild-type levels of transcription in vivo. Here, we show that poly(dA:dT) is a novel promoter element whose function depends on its intrinsic structure, not its interaction with sequence-specific, DNA-binding proteins. First, poly(dA:dT) stimulates Gcn4-activated transcription in a manner that is length dependent and inversely related to intracellular Gcn4 levels. Second, Datin, the only known poly(dA:dT)-binding protein, behaves as a repressor through poly(dA:dT) sequences. Third, poly(dG:dC), a structurally dissimilar homopolymer that also affects nucleosomes, has transcriptional properties virtually identical to those of poly(dA:dT). Three probes of chromatin structure including HinfI endonuclease cleavage in vivo indicate that poly(dA:dT) increases accessibility of the Gcn4 binding site and adjacent sequences in physiological chromatin. These observations suggest that, by virtue of its intrinsic structure, poly(dA:dT) locally affects nucleosomes and increases the accessibility of transcription factors bound to nearby sequences.
许多酵母启动子包含同聚dA:dT序列,这些序列在体外影响核小体形成,并且是体内野生型转录水平所必需的。在此,我们表明聚(dA:dT)是一种新型启动子元件,其功能取决于其固有结构,而非与序列特异性DNA结合蛋白的相互作用。首先,聚(dA:dT)以长度依赖且与细胞内Gcn4水平呈负相关的方式刺激Gcn4激活的转录。其次,已知唯一的聚(dA:dT)结合蛋白Datin通过聚(dA:dT)序列发挥阻遏作用。第三,聚(dG:dC),一种结构不同但同样影响核小体的同聚物,其转录特性与聚(dA:dT)几乎相同。包括体内HinfI内切酶切割在内的三种染色质结构探针表明,聚(dA:dT)增加了生理染色质中Gcn4结合位点及相邻序列的可及性。这些观察结果表明,聚(dA:dT)凭借其固有结构在局部影响核小体,并增加与附近序列结合的转录因子的可及性。