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番茄丁香假单胞菌 DC3000 中 Fur 调控子的特性研究。

Characterization of the Fur regulon in Pseudomonas syringae pv. tomato DC3000.

机构信息

Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, New York 14853, USA.

出版信息

J Bacteriol. 2011 Sep;193(18):4598-611. doi: 10.1128/JB.00340-11. Epub 2011 Jul 22.

Abstract

The plant pathogen Pseudomonas syringae pv. tomato DC3000 (DC3000) is found in a wide variety of environments and must monitor and respond to various environmental signals such as the availability of iron, an essential element for bacterial growth. An important regulator of iron homeostasis is Fur (ferric uptake regulator), and here we present the first study of the Fur regulon in DC3000. Using chromatin immunoprecipitation followed by massively parallel sequencing (ChIP-seq), 312 chromosomal regions were highly enriched by coimmunoprecipitation with a C-terminally tagged Fur protein. Integration of these data with previous microarray and global transcriptome analyses allowed us to expand the putative DC3000 Fur regulon to include genes both repressed and activated in the presence of bioavailable iron. Using nonradioactive DNase I footprinting, we confirmed Fur binding in 41 regions, including upstream of 11 iron-repressed genes and the iron-activated genes encoding two bacterioferritins (PSPTO_0653 and PSPTO_4160), a ParA protein (PSPTO_0855), and a two-component system (TCS) (PSPTO_3382 to PSPTO_3380).

摘要

植物病原菌丁香假单胞菌 pv. 番茄 DC3000(DC3000)存在于多种环境中,必须监测和响应各种环境信号,如铁的可用性,铁是细菌生长的必需元素。铁稳态的一个重要调节剂是 Fur(铁摄取调节剂),在这里我们首次研究了 DC3000 中的 Fur 调控子。使用染色质免疫沉淀 followed by massively parallel sequencing(ChIP-seq),通过与 C 末端标记的 Fur 蛋白的共免疫沉淀,高度富集了 312 个染色体区域。将这些数据与以前的微阵列和全转录组分析整合在一起,使我们能够将推定的 DC3000 Fur 调控子扩展到包括在生物可利用铁存在下受抑制和激活的基因。使用非放射性 DNase I 足迹分析,我们在 41 个区域中证实了 Fur 的结合,包括 11 个铁受抑制基因和编码两个细菌铁蛋白(PSPTO_0653 和 PSPTO_4160)、一个 ParA 蛋白(PSPTO_0855)和一个双组分系统(TCS)(PSPTO_3382 至 PSPTO_3380)的上游。

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