Characterization of SAV7471, a TetR-Family Transcriptional Regulator Involved in the Regulation of Coenzyme A Metabolism in Streptomyces avermitilis
Characterization of SAV7471, a TetR-Family Transcriptional Regulator Involved in the Regulation of Coenzyme A Metabolism in Streptomyces avermitilis
复制标题
SAV7471（参与阿维链霉菌辅酶 A 代谢调节的 TetR 家族转录调节因子）的表征
DOI：
10.1128/jb.00716-13
复制
发表时间：
2013-10-01
影响因子：
3.2
通讯作者：
Li, Jilun
中科院分区：
文献类型：
--
作者：
Liu, Yanping;Yan, Tingting;Li, Jilun
通过基因缺失、互补和过表达实验，研究了革兰氏阳性土壤菌阿维菌链霉菌（Streptomyces avermitilis）的四tr转录调控基因SAV7471在阿维菌素生产中的作用。SAV7471开放阅读框基因缺失导致阿维菌素过量产生。这种缺失还导致SAV7472-SAV7473转录本过表达，该转录本编码一种功能未知的蛋白和一种可能参与泛酸和辅酶a （CoA）代谢的黄蛋白。emsa和足迹分析表明，SAV7471可以与两个基因间区高度相似的回文序列结合，该区域包含5 ' cDNA末端快速扩增（5 ' -RACE）检测到的每个基因的明显转录起始位点。除了SAV7472-SAV7473外，至少有两个参与CoA代谢的基因（SAV1104和SAV1258）受到SAV7471的负调控。SAV7471通过负向调控靶基因SAV7472-SAV7473及其他参与辅酶a代谢的基因的转录，影响细胞代谢通量，从而间接调控阿维菌素的生物合成。
ABSTRACT The role of a tetR transcriptional regulatory gene (SAV7471) in avermectin production in the Gram-positive soil bacterium Streptomyces avermitilis was investigated by gene deletion, complementation, and overexpression experiments. Gene deletion of the SAV7471 open reading frame resulted in avermectin overproduction. The deletion also resulted in overexpression of SAV7472-SAV7473 transcripts, which encode a protein of unknown function and a flavoprotein possibly involved in pantothenate and coenzyme A (CoA) metabolism. EMSAs and footprinting assays showed that SAV7471 can bind to two palindromic sequences with high similarity in the intergenic region between SAV7471 and SAV7472, a region that contains the apparent transcription start sites for each gene detected by rapid amplification of 5′ cDNA ends (5′-RACE). In addition to SAV7472-SAV7473, at least two genes (SAV1104 and SAV1258) involved in CoA metabolism are negatively controlled by SAV7471. By negatively regulating the transcription of the target genes SAV7472-SAV7473 and other genes involved in CoA metabolism, SAV7471 may affect cellular metabolic flux and may thereby indirectly regulate avermectin biosynthesis.