Graduate School of Science and Engineering, Saitama University, Saitama, 338-8570, Japan.
Biological Science Research, KAO Corporation, Wakayama, 640-8580, Japan.
Plant J. 2017 Oct;92(2):189-198. doi: 10.1111/tpj.13644. Epub 2017 Aug 21.
Specific transcription factors have been identified in various heterotrophic bacterial species that regulate the sets of genes required for fatty acid metabolism. Here, we report that expression of the fab genes, encoding fatty acid biosynthetic enzymes, is regulated by the global regulator LexA in the photoautotrophic cyanobacterium Synechocystis sp. PCC 6803. Sll1626, an ortholog of the well-known LexA repressor involved in the SOS response in heterotrophic bacteria, was isolated from crude extracts of Synechocystis by DNA affinity chromatography, reflecting its binding to the upstream region of the acpP-fabF and fabI genes. An electrophoresis mobility shift assay revealed that the recombinant LexA protein can bind to the upstream region of each fab gene tested (fabD, fabH, fabF, fabG, fabZ and fabI). Quantitative RT-PCR analysis of the wild type and a lexA-disrupted mutant strain suggested that LexA acts as a repressor of the fab genes involved in initiation of fatty acid biosynthesis (fabD, fabH and fabF) and the first reductive step in the subsequent elongation cycle (fabG) under normal growth conditions. Under nitrogen-depleted conditions, downregulation of fab gene expression is partly achieved through an increase in LexA-repressing activity. In contrast, under phosphate-depleted conditions, fab gene expression is upregulated, probably due to the loss of repression by LexA. We further demonstrate that elimination of LexA largely increases the production of fatty acids in strains modified to secrete free fatty acids.
已在各种异养细菌物种中鉴定出特定的转录因子,这些因子调节脂肪酸代谢所需的基因集。在这里,我们报告说,编码脂肪酸生物合成酶的 fab 基因的表达受光自养蓝藻 Synechocystis sp. PCC 6803 中的全局调节剂 LexA 调节。Sll1626 是参与异养细菌 SOS 反应的众所周知的 LexA 抑制剂的同源物,通过 DNA 亲和层析从 Synechocystis 的粗提取物中分离出来,反映了它与 acpP-fabF 和 fabI 基因上游区域的结合。电泳迁移率变动分析显示,重组 LexA 蛋白可以与测试的每个 fab 基因(fabD、fabH、fabF、fabG、fabZ 和 fabI)的上游区域结合。野生型和 lexA 缺失突变体菌株的定量 RT-PCR 分析表明,LexA 作为脂肪酸生物合成起始(fabD、fabH 和 fabF)和随后延伸循环中第一个还原步骤(fabG)中 fab 基因的抑制剂在正常生长条件下起作用。在氮饥饿条件下,fab 基因表达的下调部分通过 LexA 抑制活性的增加来实现。相比之下,在缺磷条件下,fab 基因的表达上调,可能是由于 LexA 失去抑制作用。我们进一步证明,消除 LexA 会大大增加修饰为分泌游离脂肪酸的菌株中脂肪酸的产生。