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CRISPRi 敲低 cyabrB1 基因诱导蓝藻集胞藻 PCC 6803 中碳代谢相关的反向转录 icfG 和 sll1783 操纵子。

CRISPRi knockdown of the cyabrB1 gene induces the divergently transcribed icfG and sll1783 operons related to carbon metabolism in the cyanobacterium Synechocystis sp. PCC 6803.

机构信息

Graduate School of Science and Engineering, Saitama University.

Department of Biochemistry and Molecular Biology, Faculty of Science, Saitama University.

出版信息

J Gen Appl Microbiol. 2024 Jul 20;70(1). doi: 10.2323/jgam.2024.01.001. Epub 2024 Jan 27.

Abstract

Most cyanobacterial genomes possess more than two copies of genes encoding cyAbrBs (cyanobacterial AbrB-like proteins) having an AbrB-like DNA-binding domain at their C-terminal region. Accumulating data suggest that a wide variety of metabolic and physiologic processes are regulated by cyAbrBs. In this study, we investigated the function of the essential gene cyabrB1 (sll0359) in Synechocystis sp. PCC 6803 by using CRISPR interference technology. The conditional knockdown of cyabrB1 caused increases of cyAbrB2 transcript and protein levels. However, the effect of cyabrB1 knockdown on global gene expression profile was quite limited compared to the previously reported profound effect of knockout of cyabrB2. Among 24 up-regulated genes, 16 genes were members of the divergently transcribed icfG and sll1783 operons related to carbon metabolism. The results of this and previous studies indicate the different contributions of two cyAbrBs to transcriptional regulation of genes related to carbon, hydrogen and nitrogen metabolism. Possession of a pair of cyAbrBs has been highly conserved during the course of evolution of the cyanobacterial phylum, suggesting physiological significance of transcriptional regulation attained by their interaction.

摘要

大多数蓝藻基因组拥有两个以上的基因拷贝,这些基因编码具有 AbrB 样 DNA 结合域的 cyAbrBs(蓝藻 AbrB 样蛋白),位于其 C 末端区域。越来越多的数据表明,cyAbrBs 可以调节各种代谢和生理过程。在这项研究中,我们通过使用 CRISPR 干扰技术研究了 Synechocystis sp. PCC 6803 中的必需基因 cyabrB1(sll0359)的功能。cyabrB1 的条件敲低导致 cyAbrB2 转录本和蛋白质水平的增加。然而,与先前报道的 cyabrB2 敲除的深远影响相比,cyabrB1 敲低对全局基因表达谱的影响相当有限。在 24 个上调基因中,有 16 个基因是与碳代谢相关的反向转录 icfG 和 sll1783 操纵子的成员。本研究和以前的研究结果表明,两个 cyAbrBs 对与碳、氢和氮代谢相关基因的转录调控有不同的贡献。在蓝藻门的进化过程中,一对 cyAbrBs 的存在高度保守,这表明它们相互作用所达到的转录调控具有生理意义。

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