Müller J
MRC Laboratory of Molecular Biology, Cambridge, UK.
EMBO J. 1995 Mar 15;14(6):1209-20. doi: 10.1002/j.1460-2075.1995.tb07104.x.
Polycomb group (Pc-G) proteins act to keep homeotic genes stably and heritably silenced during Drosophila development. Here, it is shown that Polycomb (Pc), one of the Pc-G proteins, acts as a transcriptional silencer in Drosophila embryos if tethered to reporter genes by the DNA binding domain of GAL4 (i.e. as a GAL-Pc fusion protein). The results suggest that silencing by GAL-Pc requires the C-terminal portion of Pc, but not the chromodomain. If a pulse of Gal-Pc is provided, synthetic reporter genes are repressed, though only transiently. In contrast, reporter genes containing homeotic gene sequences remain stably and heritably silenced in a Pc-G gene-dependent fashion, even when GAL-Pc is no longer present. This implies that GAL-Pc recruits Pc-G proteins to DNA and suggests that maintenance of silencing requires the anchoring of Pc-G proteins to specific cis-regulatory sequences present in homeotic genes. The extent of DNA over which the Pc-G machinery acts is quite selective, as silencing established on one enhancer does not necessarily 'spread' to a juxtaposed synthetic enhancer.
多梳蛋白家族(Pc-G)蛋白在果蝇发育过程中使同源异型基因稳定且可遗传地沉默。本文表明,多梳蛋白家族中的一种蛋白多梳蛋白(Pc),如果通过GAL4的DNA结合结构域与报告基因相连(即作为GAL-Pc融合蛋白),在果蝇胚胎中可作为转录沉默因子。结果表明,GAL-Pc介导的沉默作用需要Pc的C末端部分,但不需要染色质结构域。如果提供GAL-Pc脉冲,合成报告基因会被抑制,不过只是短暂抑制。相比之下,含有同源异型基因序列的报告基因即使在不再存在GAL-Pc时,仍以Pc-G基因依赖的方式保持稳定且可遗传的沉默。这意味着GAL-Pc将Pc-G蛋白招募至DNA,并表明沉默状态的维持需要将Pc-G蛋白锚定到同源异型基因中存在的特定顺式调控序列上。Pc-G机制作用的DNA范围具有相当的选择性,因为在一个增强子上建立的沉默不一定会“扩散”到相邻的合成增强子上。