Vazquez J, Schedl P
Department of Biochemistry and Biophysics, University of California, San Francisco, California 94143, USA.
Genetics. 2000 Jul;155(3):1297-311. doi: 10.1093/genetics/155.3.1297.
Eukaryotic chromosomes are thought to be subdivided into a series of structurally and functionally independent units. Critical to this hypothesis is the identification of insulator or boundary elements that delimit chromosomal domains. The properties of a Notch mutation, facet-strawberry (fa(swb)), suggest that this small deletion disrupts such a boundary element. fa(swb) is located in the interband separating polytene band 3C7, which contains Notch, from the distal band 3C6. The fa(swb) mutation alters the structural organization of the chromosome by deleting the interband and fusing 3C7 with 3C6. Genetic studies also suggest that fa(swb) compromises the functional autonomy of Notch by allowing the locus to become sensitive to chromosomal position effects emanating from distal sequences. In the studies reported here, we show that a DNA fragment spanning the fa(swb) region can insulate reporter transgenes against chromosomal position effects and can block enhancer-promoter interactions. Moreover, we find that insulating activity is dependent on sequences deleted in fa(swb). These results provide evidence that the element defined by the fa(swb) mutation corresponds to an insulator.
真核生物染色体被认为可细分为一系列结构和功能上独立的单元。这一假说的关键在于鉴定界定染色体结构域的绝缘子或边界元件。Notch突变体facet-strawberry(fa(swb))的特性表明,这种小缺失破坏了这样一个边界元件。fa(swb)位于多线带3C7(包含Notch)与远端带3C6之间的间带中。fa(swb)突变通过缺失间带并将3C7与3C6融合,改变了染色体的结构组织。遗传学研究还表明,fa(swb)使Notch基因座对来自远端序列的染色体位置效应变得敏感,从而损害了Notch的功能自主性。在本文报道的研究中,我们表明跨越fa(swb)区域的一个DNA片段能够使报告转基因免受染色体位置效应的影响,并能阻断增强子-启动子相互作用。此外,我们发现绝缘活性依赖于fa(swb)中缺失的序列。这些结果提供了证据,证明由fa(swb)突变定义的元件对应于一个绝缘子。