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在游动放线菌SE50/110中,一个受麦芽糖调控的大基因组区域被转录调节因子MalT激活。

A maltose-regulated large genomic region is activated by the transcriptional regulator MalT in Actinoplanes sp. SE50/110.

作者信息

Droste Julian, Kulisch Martin, Wolf Timo, Schaffert Lena, Schneiker-Bekel Susanne, Pühler Alfred, Kalinowski Jörn

机构信息

Microbial Genomics and Biotechnology, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615, Bielefeld, Germany.

Senior Research Group in Genome Research of Industrial Microorganisms, Center for Biotechnology, Bielefeld University, Universitätsstraße 27, 33615, Bielefeld, Germany.

出版信息

Appl Microbiol Biotechnol. 2020 Nov;104(21):9283-9294. doi: 10.1007/s00253-020-10923-2. Epub 2020 Sep 28.

Abstract

Actinoplanes sp. SE50/110 is the industrially relevant producer of acarbose, which is used in the treatment of diabetes mellitus. Recent studies elucidated the expression dynamics in Actinoplanes sp. SE50/110 during growth. From these data, we obtained a large genomic region (ACSP50_3900 to ACSP50_3950) containing 51 genes, of which 39 are transcribed in the same manner. These co-regulated genes were found to be stronger transcribed on maltose compared with glucose as a carbon source. The transcriptional regulator MalT was identified as an activator of this maltose-regulated large genomic region (MRLGR). Since most of the genes are poorly annotated, the function of this region is farther unclear. However, comprehensive BLAST analyses indicate similarities to enzymes involved in amino acid metabolism. We determined a conserved binding motif of MalT overlapping the -35 promoter region of 17 transcription start sites inside the MRLGR. The corresponding sequence motif 5'-TCATCC-5nt-GGATGA-3' displays high similarities to reported MalT binding sites in Escherichia coli and Klebsiella pneumoniae, in which MalT is the activator of mal genes. A malT deletion and an overexpression mutant were constructed. Differential transcriptome analyses revealed an activating effect of MalT on 40 of the 51 genes. Surprisingly, no gene of the maltose metabolism is affected. In contrast to many other bacteria, MalT is not the activator of mal genes in Actinoplanes sp. SE50/110. Finally, the MRLGR was found partly in other closely related bacteria of the family Micromonosporaceae. Even the conserved MalT binding site was found upstream of several genes inside of the corresponding regions. KEY POINTS : • MalT is the maltose-dependent activator of a large genomic region in ACSP50_WT. • The consensus binding motif is similar to MalT binding sites in other bacteria. • MalT is not the regulator of genes involved in maltose metabolism in ACSP50_WT.

摘要

游动放线菌SE50/110是阿卡波糖的工业相关生产菌,阿卡波糖用于治疗糖尿病。最近的研究阐明了游动放线菌SE50/110在生长过程中的表达动态。从这些数据中,我们获得了一个包含51个基因的大基因组区域(ACSP50_3900至ACSP50_3950),其中39个基因以相同方式转录。与作为碳源的葡萄糖相比,发现这些共调控基因在麦芽糖上转录更强。转录调节因子MalT被鉴定为这个麦芽糖调节的大基因组区域(MRLGR)的激活剂。由于大多数基因注释不佳,该区域的功能更不清楚。然而,全面的BLAST分析表明与参与氨基酸代谢的酶有相似性。我们确定了MalT的一个保守结合基序,它与MRLGR内17个转录起始位点的-35启动子区域重叠。相应的序列基序5'-TCATCC-5nt-GGATGA-3'与大肠杆菌和肺炎克雷伯菌中报道的MalT结合位点高度相似,其中MalT是mal基因的激活剂。构建了一个malT缺失突变体和一个过表达突变体。差异转录组分析揭示了MalT对51个基因中的40个基因有激活作用。令人惊讶的是,麦芽糖代谢的基因没有受到影响。与许多其他细菌不同,MalT不是游动放线菌SE50/110中mal基因的激活剂。最后,在小单孢菌科的其他密切相关细菌中部分发现了MRLGR。甚至在相应区域内几个基因的上游也发现了保守的MalT结合位点。要点:•MalT是ACSP50_WT中一个大基因组区域的麦芽糖依赖性激活剂。•共有结合基序与其他细菌中的MalT结合位点相似。•MalT不是ACSP50_WT中参与麦芽糖代谢的基因的调节因子。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f7f/7567727/0c3498ba5f5e/253_2020_10923_Fig1_HTML.jpg

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