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.

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Atsuko Hishida, Ryo Shirai, Akiyoshi Higo, Minenosuke Matsutani, Kaori Nimura-Matsune, Tomoko Takahashi, Satoru Watanabe, Shigeki Ehira, Yukako Hihara
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引用次数: 0

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.

CRISPRi 敲除 cyabrB1 基因可诱导蓝藻 Synechocystis sp. PCC 6803 中与碳代谢相关的不同转录的 icfG 和 sll1783 操作子。
大多数蓝藻基因组都有两个以上的编码 cyAbrBs(蓝藻 AbrB 样蛋白)的基因拷贝,这些基因的 C 端区域具有 AbrB 样 DNA 结合域。不断积累的数据表明,cyAbrBs 可调控多种代谢和生理过程。在本研究中,我们利用 CRISPR 干扰技术研究了 Synechocystis sp.条件性敲除cyabrB1会导致cyAbrB2转录本和蛋白水平的升高。然而,与之前报道的敲除 cyabrB2 的深远影响相比,敲除 cyabrB1 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 个基因属于与碳代谢有关的不同转录的 icfG 和 sll1783 操作子。这项研究和以前的研究结果表明,两个 cyAbrB 对碳、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,拥有一对 cyAbrBs 是高度保守的,这表明通过它们的相互作用实现转录调控具有重要的生理意义。
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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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