RedB, a Member of the CRP/FNR Family, Functions as a Transcriptional Redox Brake

• Published in: Microbiology spectrum 2022-10, Vol.10 (5), p.e0235322-e0235322
• Main Authors: Ke, Nijia, Kumka, Joseph E, Fang, Mingxu, Weaver, Brian, Burstyn, Judith N, Bauer, Carl E
• Format: Article
• Language: English
• Subjects: Adenosine Triphosphate - metabolism; Amino Acids - metabolism; Bacterial Proteins - genetics; Bacterial Proteins - metabolism; CRP/FNR ortholog; Cyclic AMP Receptor Protein - genetics; Cyclic AMP Receptor Protein - metabolism; Escherichia coli Proteins - metabolism; Fumarates; Gene Expression Regulation, Bacterial; Genetics and Molecular Biology; global regulation; Iron-Sulfur Proteins - metabolism; limiting energy production; NAD - genetics; NAD - metabolism; Oxidation-Reduction; Phylogeny; Research Article; RNA, Transfer - metabolism; Transcription Factors - metabolism; transcriptomics; Tricarboxylic Acids
• ISSN: 2165-0497; 2165-0497
• DOI: 10.1128/spectrum.02353-22

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.