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新型 insight 揭示了蓝藻集胞藻 PCC 6803 中 LexA 的调控机制。

Novel insights into the regulation of LexA in the cyanobacterium Synechocystis sp. Strain PCC 6803.

机构信息

Department of Photochemistry and Molecular Science, Ångström Laboratories, Uppsala University, Uppsala, Sweden.

出版信息

J Bacteriol. 2011 Aug;193(15):3804-14. doi: 10.1128/JB.00289-11. Epub 2011 Jun 3.

Abstract

The transcription factor LexA in the cyanobacterium Synechocystis sp. strain PCC 6803 has been shown to regulate genes that are not directly involved in DNA repair but instead in several different metabolic pathways. However, the signal transduction pathways remain largely uncharacterized. The present work gives novel insights into the regulation of LexA in this unicellular cyanobacterium. A combination of Northern and Western blotting, using specific antibodies against the cyanobacterial LexA, was employed to show that this transcription regulator is under posttranscriptional control, in addition to the classical and already-described transcriptional regulation. Moreover, detailed two-dimensional (2D) electrophoresis analyses of the protein revealed that LexA undergoes posttranslational modifications. Finally, a fully segregated LexA::GFP (green fluorescent protein) fusion-modified strain was produced to image LexA's spatial distribution in live cells. The fusion protein retains DNA binding capabilities, and the GFP fluorescence indicates that LexA is localized in the innermost region of the cytoplasm, decorating the DNA in an evenly distributed pattern. The implications of these findings for the overall role of LexA in Synechocystis sp. strain PCC 6803 are further discussed.

摘要

已证实,蓝藻集胞藻 PCC 6803 中的转录因子 LexA 可调控那些不直接参与 DNA 修复,但却参与多种不同代谢途径的基因。然而,其信号转导途径在很大程度上仍未被描述。本研究工作为该单细胞蓝藻中 LexA 的调控提供了新的见解。采用Northern 和 Western 印迹的组合,使用针对蓝细菌 LexA 的特异性抗体,表明该转录调控因子不仅受到经典的、已经描述过的转录调控,还受到转录后调控的影响。此外,对蛋白质的详细二维(2D)电泳分析表明 LexA 经历了翻译后修饰。最后,构建了完全分隔的 LexA::GFP(绿色荧光蛋白)融合修饰菌株,以在活细胞中对 LexA 的空间分布进行成像。融合蛋白保留了 DNA 结合能力,GFP 荧光表明 LexA 定位于细胞质的最内层区域,以均匀分布的模式装饰 DNA。这些发现对 LexA 在集胞藻 PCC 6803 中的整体作用的影响将进一步讨论。

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Computational analysis of LexA regulons in Cyanobacteria.蓝藻 LexA 调控子的计算分析。
BMC Genomics. 2010 Sep 29;11:527. doi: 10.1186/1471-2164-11-527.

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