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果蝇中多梳蛋白组染色质复合物成分的募集

Recruitment of components of Polycomb Group chromatin complexes in Drosophila.

作者信息

Poux S, McCabe D, Pirrotta V

机构信息

Department of Zoology, University of Geneva, CH1211 Geneva, Switzerland.

出版信息

Development. 2001 Jan;128(1):75-85. doi: 10.1242/dev.128.1.75.

Abstract

Polycomb Group complexes assemble at polycomb response elements (PREs) in vivo and silence genes in the surrounding chromatin. To study the recruitment of silencing complexes, we have targeted various Polycomb Group (PcG) proteins by fusing them to the LexA DNA binding domain. When LexA-PC, -PSC, -PH or -SU(Z)2 are targeted to a reporter gene, they recruit functional PcG-silencing complexes that recapitulate the silencing behavior of a PRE: silencing is sensitive to the state of activity of the target chromatin. When the target is transcriptionally active, silencing is not established but when the target is not active at syncytial blastoderm, it becomes silenced. The repressed state persists through embryonic development but cannot be maintained in larval imaginal discs even when the LexA-PcG fusion is constitutively expressed, suggesting a discontinuity in the mechanism of repression. These proteins also interact with other PC-containing complexes in embryonic nuclear extracts. In contrast LexA-PHO is neither able to silence nor to interact with PC-containing complexes. Analysis of pho mutant embryos and of PRE constructs whose PHO-binding sites are mutated suggests that, while PHO is important for silencing in imaginal discs, it is not necessary for embryonic PcG silencing.

摘要

多梳蛋白复合体在体内多梳反应元件(PREs)处组装,并使周围染色质中的基因沉默。为了研究沉默复合体的招募过程,我们通过将各种多梳蛋白复合体(PcG)与LexA DNA结合结构域融合来靶向它们。当LexA-PC、-PSC、-PH或-SU(Z)2靶向报告基因时,它们会招募功能性的PcG沉默复合体,这些复合体重现了PRE的沉默行为:沉默对靶染色质的活性状态敏感。当靶标处于转录活性状态时,沉默不会建立,但当靶标在合胞体胚盘期不活跃时,它就会被沉默。这种抑制状态在胚胎发育过程中持续存在,但即使LexA-PcG融合蛋白持续表达,在幼虫的成虫盘也无法维持,这表明抑制机制存在间断性。这些蛋白还能与胚胎核提取物中其他含PC的复合体相互作用。相比之下,LexA-PHO既不能沉默,也不能与含PC的复合体相互作用。对pho突变胚胎和PHO结合位点发生突变的PRE构建体的分析表明,虽然PHO对成虫盘中的沉默很重要,但对胚胎期PcG沉默并非必需。

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