Molecular and Cellular Biochemistry Department, Indiana University, Bloomington, Indiana, USA.
Department of Chemistry, University of Wisconsin-Madison, Madison, Wisconsin, USA.
Microbiol Spectr. 2022 Oct 26;10(5):e0235322. doi: 10.1128/spectrum.02353-22. Epub 2022 Sep 15.
Phylogenetic and sequence similarity network analyses of the CRP (cyclic AMP receptor protein)/FNR (fumarate and nitrate reductase regulatory protein) family of transcription factors indicate the presence of numerous subgroups, many of which have not been analyzed. Five homologs of the CRP/FNR family are present in the Rhodobacter capsulatus genome. One is a member of a broadly disseminated, previously uncharacterized CRP/FNR family subgroup encoded by the gene . In this study, we utilize mutational disruption, transcriptome sequencing (RNA-seq), and chromatin immunoprecipitation sequencing (ChIP-seq) to determine the role of RCC01561 in regulating R. capsulatus physiology. This analysis shows that a mutant strain disrupted for exhibits altered expression of 451 genes anaerobically. A detailed analysis of the affected loci shows that RCC01561 represses photosynthesis and favors catabolism over anabolism and the use of the Entner-Doudoroff shunt and glycolysis over that of the tricarboxylic acid (TCA) cycle to limit NADH and ATP formation. This newly characterized CRP/FNR family member with a predominant role in reducing the production of reducing potential and ATP is given the nomenclature RedB as it functions as an energy and ox rake. Beyond limiting energy production, RedB also represses the expression of numerous genes involved in protein synthesis, including those involved in translation initiation, tRNA synthesis and charging, and amino acid biosynthesis. CRP and FNR are well-characterized members of the CRP/FNR family of regulatory proteins that function to maximize cellular energy production. In this study, we identify several new subgroups of the CRP/FNR family, many of which have not yet been characterized. Using Rhodobacter capsulatus as a model, we have mutationally disrupted the gene , which codes for a transcription factor that is a member of a unique subgroup of the CRP/FNR family. Transcriptomic analysis shows that the disruption of leads to the altered expression of 451 genes anaerobically. Analysis of these regulated genes indicates that RCC01561 has a novel role in limiting cellular energy production. To our knowledge, this is first example of a member of the CRP/FNR family that functions as a brake on cellular energy production.
CRP(环腺苷酸受体蛋白)/FNR(延胡索酸和硝酸盐还原酶调节蛋白)家族转录因子的系统发育和序列相似性网络分析表明,存在许多亚群,其中许多尚未进行分析。在荚膜红细菌基因组中有 5 个 CRP/FNR 家族的同源物。其中一个是由基因编码的广泛传播的、以前未被描述的 CRP/FNR 家族亚群的成员。在本研究中,我们利用突变破坏、转录组测序(RNA-seq)和染色质免疫沉淀测序(ChIP-seq)来确定 RCC01561 在调节荚膜红细菌生理学中的作用。该分析表明,一个破坏的突变菌株在厌氧条件下表现出 451 个基因的表达改变。对受影响基因座的详细分析表明,RCC01561 抑制光合作用,有利于分解代谢而不是合成代谢,并且有利于使用 Entner-Doudoroff 旁路和糖酵解而不是三羧酸(TCA)循环来限制 NADH 和 ATP 的形成。这个新鉴定的 CRP/FNR 家族成员主要作用是降低还原潜力和 ATP 的产生,被命名为 RedB,因为它作为能量和氧化耙子发挥作用。除了限制能量产生外,RedB 还抑制了许多参与蛋白质合成的基因的表达,包括那些参与翻译起始、tRNA 合成和加载以及氨基酸生物合成的基因。CRP 和 FNR 是 CRP/FNR 家族调节蛋白的典型成员,其功能是最大限度地提高细胞的能量产生。在本研究中,我们鉴定了 CRP/FNR 家族的几个新亚群,其中许多尚未被描述。我们以荚膜红细菌为模型,突变破坏了编码转录因子的基因,该转录因子是 CRP/FNR 家族的一个独特亚群的成员。转录组分析表明,破坏导致 451 个基因在厌氧条件下的表达改变。对这些受调控基因的分析表明,RCC01561 在限制细胞能量产生方面具有新的作用。据我们所知,这是 CRP/FNR 家族成员作为细胞能量产生制动器的第一个例子。