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盘基网柄菌bZIP转录因子DimB调节前柄特异性基因表达。

The Dictyostelium bZIP transcription factor DimB regulates prestalk-specific gene expression.

作者信息

Zhukovskaya Natasha V, Fukuzawa Masashi, Yamada Yoko, Araki Tsuyoshi, Williams Jeffrey G

机构信息

School of Life Sciences, University of Dundee, MSI/WTB Complex, UK.

出版信息

Development. 2006 Feb;133(3):439-48. doi: 10.1242/dev.02190. Epub 2006 Jan 5.

Abstract

The ecmA gene is specifically expressed in prestalk cells and its transcription is induced by the chlorinated hexaphenone DIF-1. We have purified a novel bZIP transcription factor, DimB, by affinity chromatography on two spatially separated ecmA promoter fragments. Mutagenesis of the cap-site proximal DimB-binding site (the -510 site) greatly decreases ecmA expression in the pstO cells, which comprise the rear half of the prestalk zone, and also in the Anterior-Like Cells, which lie scattered throughout the prespore region. However, DimB is not essential for normal expression of the ecmA gene, instead it spatially limits its expression; ecmA is relatively highly expressed in the subset of prestalk cells that coats the prestalk zone, but in slugs of a DimB-null strain, ecmA is highly expressed throughout the prestalk zone. Because the -510 site is required for correct ecmA expression, we posit a separate activator protein that competes with DimB for binding to the -510 site. DimB rapidly accumulates in the nucleus when cells are exposed to DIF-1, and ChIP analysis shows that, in the presence of extracellular cAMP, DIF-1 causes DimB to associate with the ecmA promoter in vivo. Thus, DIF-1 regulates DimB activity to generate a gradient of ecmA expression in the prestalk zone of the slug.

摘要

ecmA基因在柄细胞前体细胞中特异性表达,其转录由氯化六苯酮DIF-1诱导。我们通过在两个空间分离的ecmA启动子片段上进行亲和层析,纯化了一种新型的bZIP转录因子DimB。帽位点近端的DimB结合位点(-510位点)的诱变极大地降低了ecmA在pstO细胞中的表达,pstO细胞构成柄细胞前体区域的后半部分,也降低了在类似前部细胞中的表达,这些细胞散布在整个孢子前区域。然而,DimB对于ecmA基因的正常表达并非必需,相反,它在空间上限制了其表达;ecmA在覆盖柄细胞前体区域的柄细胞前体细胞亚群中相对高度表达,但在DimB缺失菌株的蛞蝓中,ecmA在整个柄细胞前体区域都高度表达。由于-510位点对于ecmA的正确表达是必需的,我们推测存在一种与DimB竞争结合-510位点的单独激活蛋白。当细胞暴露于DIF-1时,DimB会迅速在细胞核中积累,染色质免疫沉淀分析表明,在细胞外cAMP存在的情况下,DIF-1会导致DimB在体内与ecmA启动子结合。因此,DIF-1调节DimB的活性,以在蛞蝓的柄细胞前体区域产生ecmA表达梯度。

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