Zink D, Paro R
ZMBH, Center for Molecular Biology, University of Heidelberg, Germany.
EMBO J. 1995 Nov 15;14(22):5660-71. doi: 10.1002/j.1460-2075.1995.tb00253.x.
The genes of the Polycomb-group (Pc-G) are responsible for maintaining the inactive expression state of homeotic genes. They act through specific cis-regulatory DNA elements termed PREs (Pc-G Response Elements). Multimeric complexes containing the Pc-G proteins are thought to induce heterochromatin-like structures, which stably and heritably inactivate transcription. We have tested the functional role of the FAB fragment, a PRE of the bithorax complex. We find that this element behaves as an orientation dependent silencer, capable of inducing mosaic gene expression on neighboring genes. Transgenic fly lines were constructed containing a PRE adjacent to a reporter gene inducible by the yeast GAL4 trans-activator. The competition between the activator and Pc-G-containing chromatin was visualized on polytene chromosomes using immunocytochemistry. The Pc-G protein Polycomb and GAL4 have mutually exclusive binding patterns, supporting the notion that Pc-G-induced chromatin structures can prevent activators from binding to their target sequences. However, this antagonistic function can be overcome by high doses of GAL4, even in the absence of DNA replication.
多梳家族(Pc-G)基因负责维持同源异型基因的非活性表达状态。它们通过称为PREs(Pc-G反应元件)的特定顺式调控DNA元件发挥作用。含有Pc-G蛋白的多聚体复合物被认为可诱导异染色质样结构,从而稳定且可遗传地使转录失活。我们测试了双胸复合体的一个PRE——FAB片段的功能作用。我们发现该元件表现为一种方向依赖性沉默子,能够诱导相邻基因的镶嵌基因表达。构建了转基因果蝇品系,其中一个PRE与可被酵母GAL4反式激活因子诱导的报告基因相邻。利用免疫细胞化学技术在多线染色体上观察了激活因子与含Pc-G的染色质之间的竞争。Pc-G蛋白多梳和GAL4具有相互排斥的结合模式,这支持了Pc-G诱导的染色质结构可阻止激活因子与其靶序列结合的观点。然而,即使在没有DNA复制的情况下,高剂量的GAL4也能克服这种拮抗作用。