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首页> 外文期刊>Applied Microbiology >Differential RNA Sequencing Implicates Sulfide as the Master Regulator of S0 Metabolism in Chlorobaculum tepidum and Other Green Sulfur Bacteria
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Differential RNA Sequencing Implicates Sulfide as the Master Regulator of S0 Metabolism in Chlorobaculum tepidum and Other Green Sulfur Bacteria

机译：差异RNA测序涉及硫化物作为温和性绿藻和其他绿色硫细菌中S0代谢的主要调节剂。

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The green sulfur bacteria (Chlorobiaceae) are anaerobes that use electrons from reduced sulfur compounds (sulfide, S0, and thiosulfate) as electron donors for photoautotrophic growth. Chlorobaculum tepidum, the model system for the Chlorobiaceae, both produces and consumes extracellular S0 globules depending on the availability of sulfide in the environment. These physiological changes imply significant changes in gene regulation, which has been observed when sulfide is added to Cba. tepidum growing on thiosulfate. However, the underlying mechanisms driving these gene expression changes, i.e., the specific regulators and promoter elements involved, have not yet been defined. Here, differential RNA sequencing (dRNA-seq) was used to globally identify transcript start sites (TSS) that were present during growth on sulfide, biogenic S0, and thiosulfate as sole electron donors. TSS positions were used in combination with RNA-seq data from cultures growing on these same electron donors to identify both basal promoter elements and motifs associated with electron donor-dependent transcriptional regulation. These motifs were conserved across homologous Chlorobiaceae promoters. Two lines of evidence suggest that sulfide-mediated repression is the dominant regulatory mode in Cba. tepidum. First, motifs associated with genes regulated by sulfide overlap key basal promoter elements. Second, deletion of the Cba. tepidum1277 (CT1277) gene, encoding a putative regulatory protein, leads to constitutive overexpression of the sulfide:quinone oxidoreductase CT1087 in the absence of sulfide. The results suggest that sulfide is the master regulator of sulfur metabolism in Cba. tepidum and the Chlorobiaceae. Finally, the identification of basal promoter elements with differing strengths will further the development of synthetic biology in Cba. tepidum and perhaps other Chlorobiaceae.IMPORTANCE Elemental sulfur is a key intermediate in biogeochemical sulfur cycling. The photoautotrophic green sulfur bacterium Chlorobaculum tepidum either produces or consumes elemental sulfur depending on the availability of sulfide in the environment. Our results reveal transcriptional dynamics of Chlorobaculum tepidum on elemental sulfur and increase our understanding of the mechanisms of transcriptional regulation governing growth on different reduced sulfur compounds. This report identifies genes and sequence motifs that likely play significant roles in the production and consumption of elemental sulfur. Beyond this focused impact, this report paves the way for the development of synthetic biology in Chlorobaculum tepidum and other Chlorobiaceae by providing a comprehensive identification of promoter elements for control of gene expression, a key element of strain engineering.
机译：绿色的硫细菌（绿藻科）是厌氧菌，利用还原的硫化合物（硫化物，SO和硫代硫酸盐）中的电子作为光合养分生长的电子供体。绿球藻科的模型系统-绿球藻（Chlorobaculum tepidum），根据环境中硫化物的可用性，既产生也消耗细胞外的SO球。这些生理变化暗示着基因调控的显着变化，这是在将硫化物添加到Cba中时观察到的。在硫代硫酸盐上生长的温热菌。然而，尚未定义驱动这些基因表达变化的潜在机制，即所涉及的特定调节子和启动子元件。在这里，差异RNA测序（dRNA-seq）用于全局鉴定在硫化物，生物源SO和硫代硫酸盐作为唯一电子供体的生长过程中存在的转录起始位点（TSS）。将TSS位置与在这些相同电子供体上生长的培养物中的RNA-seq数据结合使用，以识别基础启动子元件和与电子供体依赖性转录调控相关的基序。这些基序在同源的蓝藻科启动子中是保守的。有两条证据表明，硫化物介导的抑制是Cba的主要调节方式。温热的。首先，与受硫化物调控的基因相关的基序与关键的基础启动子元件重叠。第二，删除Cba。 tepidum1277（CT1277）基因编码假定的调节蛋白，导致在不存在硫化物的情况下，硫化物：醌氧化还原酶CT1087的组成型过表达。结果表明，硫化物是Cba中硫代谢的主要调节剂。 tepidum和绿藻科。最后，鉴定具有不同强度的基础启动子元件将进一步促进Cba合成生物学的发展。重要元素硫是生物地球化学硫循环中的关键中间体。光能自养的绿色硫细菌温热绿藻（Chlorobaculum tepidum）根据环境中硫化物的可用性产生或消耗元素硫。我们的研究结果揭示了温热Chlorobaculum pidpidum在元素硫上的转录动力学，并增加了我们对控制不同还原硫化合物生长的转录调控机制的理解。该报告确定了可能在元素硫的产生和消耗中起重要作用的基因和序列基序。除此重点影响外，本报告还通过全面鉴定启动子元件来控制基因表达，这是菌株工程设计的关键要素，从而为温州绿藻和其他蓝藻科的合成生物学的发展铺平了道路。

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