Transcriptional Regulation of the Daptomycin Gene Cluster in Streptomyces roseosporus by an Autoregulator, AtrA
Transcriptional Regulation of the Daptomycin Gene Cluster in Streptomyces roseosporus by an Autoregulator, AtrA
复制标题
玫瑰孢链霉菌中达托霉素基因簇通过自动调节因子 AtrA 的转录调节。
DOI：
10.1074/jbc.m114.608273
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发表时间：
2015-03-20
影响因子：
4.8
通讯作者：
Li, Yong-Quan
中科院分区：
文献类型：
--
作者：
Mao, Xu-Ming;Luo, Shuai;Li, Yong-Quan
背景：目前还没有在转录调控水平上研究达托霉素生产的报道。结果：自身调节子ATRA直接调控达托霉素基因簇的表达，ATRA是AdpA的转录靶点。结论：ATRA介导的转录信号通路直接调节达托霉素的合成。意义：我们首次揭示了达托霉素生产的转录调控机制，为其潜在的合理基因工程提供了可能。达托霉素是一种由玫瑰链霉菌产生的环状脂肽抗生素。为了揭示达托霉素生物合成的转录调控机制，我们使用生物素化的dptE启动子(DptEp)作为探针，从菌丝体蛋白质组中亲和分离了TetR家族转录调节因子dptEp相互作用蛋白ATRA。ATRA直接与dptEp结合，正向调节基因簇的表达和达托霉素的产生。同时，ATRA和adpA突变体均表现为秃顶表型和达托霉素零产生。AdpA通过与ATRA启动子(ATRAP)的直接相互作用正向调节ATRA的表达，并在A因子受体的同源物玫瑰孢霉中去除ARPA，导致形态发育加速和达托霉素产量增加，表明ATRA是AdpA介导A因子信号通路的靶标。此外，ATRA通过与其自身的启动子ATRAP结合而被正向自我调节。因此，我们第一次在转录水平上确定了一个自身调节因子ATRA，它直接介导A因子信号通路来调节达托霉素的适当产生。
Background: No investigation on daptomycin production at the transcriptional regulatory level has been reported. Results: The autoregulator AtrA directly regulates daptomycin gene cluster expression, and atrA is the transcriptional target of AdpA. Conclusion: The AtrA-mediated transcriptional signaling pathway directly regulates daptomycin production. Significance: We reveal for the first time the transcriptional regulatory mechanism of daptomycin production for its potential rational genetic engineering.Daptomycin is a cyclic lipopeptide antibiotic produced by Streptomyces roseosporus. To reveal the transcriptional regulatory mechanism of daptomycin biosynthesis, we used the biotinylated dptE promoter (dptEp) as a probe to affinity isolate the dptEp-interactive protein AtrA, a TetR family transcriptional regulator, from the proteome of mycelia. AtrA bound directly to dptEp to positively regulate gene cluster expression and daptomycin production. Meanwhile, both atrA and adpA mutants showed bald phenotype and null production of daptomycin. AdpA positively regulated atrA expression by direct interaction with atrA promoter (atrAp), and removal of ArpA in S. roseosporus, a homolog of the A-factor receptor, resulted in accelerated morphological development and increased daptomycin production, suggesting that atrA was the target of AdpA to mediate the A-factor signaling pathway. Furthermore, AtrA was positively autoregulated by binding to its own promoter atrAp. Thus, for the first time at the transcriptional level, we have identified an autoregulator, AtrA, that directly mediates the A-factor signaling pathway to regulate the proper production of daptomycin.