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GcvB小RNA对芳香族氨基酸代谢的转录后调控 于……中

Post-transcriptional regulation of aromatic amino acid metabolism by GcvB small RNA in .

作者信息

Kanda Takeshi, Sekijima Toshiko, Miyakoshi Masatoshi

机构信息

Department of Infection Biology, Institute of Medicine, University of Tsukuba, Ibaraki, Japan.

Transborder Medical Research Center, University of Tsukuba, Ibaraki, Japan.

出版信息

Microbiol Spectr. 2025 Mar 4;13(3):e0203524. doi: 10.1128/spectrum.02035-24. Epub 2025 Jan 27.

Abstract

synthesizes aromatic amino acids (AAAs) through the common pathway to produce the precursor, chorismate, and the three terminal pathways to convert chorismate into Phe, Tyr, and Trp. also imports exogenous AAAs through five transporters. GcvB small RNA post-transcriptionally regulates more than 50 genes involved in amino acid uptake and biosynthesis in , but the full extent of GcvB regulon is still underestimated. This study examined all genes involved in AAA biosynthesis and transport using translation reporter assay and qRT-PCR analysis. In addition to previously verified targets, , , and , we identified new target genes that were significantly repressed by GcvB primarily via the R1 seed region. Exceptionally, GcvB strongly inhibits the expression of , which encodes the major isozyme of the first reaction in the common pathway, through direct base pairing between the translation initiation region and the GcvB R3 seed sequence. RNase E mediates the degradation of target mRNAs except and via its C-terminal domain. GcvB overexpression prolongs the lag phase and reduces the growth rate in minimal media supplemented with AAAs and confers resistance to an antibiotic compound, azaserine, by repressing AAA transporters.IMPORTANCE strains have been genetically modified in relevant transcription factors and biosynthetic enzymes for industrial use in the fermentative production of aromatic amino acids (AAAs) and their derivative compounds. This study focuses on GcvB small RNA, a global regulator of amino acid metabolism in , and identifies new GcvB targets involved in AAA biosynthesis and uptake. GcvB represses the expression of the first and last enzymes of the common pathway and the first enzymes of Trp and Phe terminal pathways. GcvB also limits import of AAAs. This paper documents the impact of RNA-mediated regulation on AAA metabolism in .

摘要

通过共同途径合成芳香族氨基酸(AAA)以产生前体分支酸,以及通过三条末端途径将分支酸转化为苯丙氨酸(Phe)、酪氨酸(Tyr)和色氨酸(Trp)。它还通过五种转运蛋白导入外源AAA。GcvB小RNA在转录后调节超过50个参与氨基酸摄取和生物合成的基因,但GcvB调控子的完整范围仍被低估。本研究使用翻译报告基因测定和qRT-PCR分析检查了所有参与AAA生物合成和转运的基因。除了先前验证的靶标、和外,我们还鉴定了主要通过R1种子区域被GcvB显著抑制的新靶基因。特别地,GcvB通过翻译起始区域与GcvB R3种子序列之间的直接碱基配对,强烈抑制编码共同途径中第一个反应的主要同工酶的表达。核糖核酸酶E通过其C末端结构域介导除和之外的靶标mRNA的降解。GcvB过表达延长了在补充有AAA的基本培养基中的延迟期并降低了生长速率,并且通过抑制AAA转运蛋白赋予对抗生素化合物重氮丝氨酸的抗性。重要性在相关转录因子和生物合成酶中对菌株进行了基因改造,以用于芳香族氨基酸(AAA)及其衍生化合物的发酵生产的工业用途。本研究聚焦于GcvB小RNA,其是氨基酸代谢的全局调节因子,并鉴定了参与AAA生物合成和摄取的新GcvB靶标。GcvB抑制共同途径的第一个和最后一个酶以及Trp和Phe末端途径的第一个酶的表达。GcvB还限制AAA的导入。本文记录了RNA介导的调节对氨基酸代谢的影响。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ff3e/11878033/13ba753f4501/spectrum.02035-24.f001.jpg

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