Protein occupancy landscape of a bacterial genome.

Protein occupancy landscape of a bacterial genome.
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DOI:
10.1016/j.molcel.2009.06.035
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发表时间:
2009-07-31
期刊:
影响因子:
16
通讯作者:
Tavazoie S
Tavazoie S
中科院分区:
生物学1区
文献类型:
--
作者:
Vora T;Hottes AK;Tavazoie S

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蛋白质-DNA相互作用是核心生物过程的基础,包括转录、DNA复制和染色体组织。我们开发了体内蛋白质占用显示(IPOD)技术,该技术可以在单个结合位点的分辨率上显示整个细菌染色体上的蛋白质占用。应用于大肠杆菌发现了数千个蛋白质占用峰,在非编码调控区域内和附近高度富集。此外,我们还发现了广泛的(bbb10千碱基)蛋白占用域(epod),其中一些定位于高表达基因,在rna聚合酶占用中富集。然而,大多数都定位于由保守的假设orf主导的转录沉默位点。这些区域高度富含预测和实验确定的类核蛋白结合位点,并表现出极高的生物物理特征,如高固有曲率。我们的观察暗示这些转录沉默的epod作为难以捉摸的组织中心,长期以来被认为是在拓扑上分离染色体结构域。
Protein-DNA interactions are fundamental to core biological processes including transcription, DNA replication, and chromosomal organization. We have developed In vivo Protein Occupancy Display (IPOD), a technology that reveals protein occupancy across an entire bacterial chromosome at the resolution of individual binding sites. Application to Escherichia coli reveals thousands of protein occupancy peaks, highly enriched within and in close proximity to non-coding regulatory regions. In addition, we discovered extensive (>1 kilobase) protein occupancy domains (EPODs), some of which are localized to highly-expressed genes, enriched in RNA-polymerase occupancy. However, the majority are localized to transcriptionally-silent loci dominated by conserved hypothetical ORFs. These regions are highly enriched in both predicted and experimentally determined binding sites of nucleoid proteins and exhibit extreme biophysical characteristics such as high intrinsic curvature. Our observations implicate these transcriptionally-silent EPODs as the elusive organizing centers, long proposed to topologically isolate chromosomal domains.
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