ENAapplication/xmlftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/034/SRR13530934/SRR13530934_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/033/SRR13530933/SRR13530933_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/032/SRR13530932/SRR13530932_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/035/SRR13530935/SRR13530935_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/037/SRR13530937/SRR13530937_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/033/SRR13530933/SRR13530933_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/034/SRR13530934/SRR13530934_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/036/SRR13530936/SRR13530936_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/037/SRR13530937/SRR13530937_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/036/SRR13530936/SRR13530936_2.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/032/SRR13530932/SRR13530932_1.fastq.gzftp://ftp.sra.ebi.ac.uk/vol1/fastq/SRR135/035/SRR13530935/SRR13530935_1.fastq.gzprimary200OKGenomicsCenter for Biotechnology, Bielefeld Universityhttps://www.ebi.ac.uk/ena/browser/view/PRJNA694916Corynebacterium diphtheriaepathogen:bacteriumxref:PubMed:33672886pathogenRNA degradation is a crucial process in bacterial cells for maintaining proper transcriptome homeostasis and coping with changing environments. A specialized ribonuclease known as RNase J (RnJ) participates in mRNA turnover in many Gram-positive bacteria however, nothing is known about this process in Corynebacterium diphtheriae, nor is the identity of this RNase. We report here that C. diphtheriae DIP1463 encodes a predicted RnJ homolog, comprised of an N-terminal beta-lactamase domain, followed by beta-CASP and C-terminal domains. We show that a recombinant protein encompassing the beta-lactamase domain possessed 5’-exoribonuclease activity, which was abolished by alanine-substitution of conserved catalytic residues His186 and His188. Intriguingly, deletion of DIP1463/rnj in C. diphtheriae caused slow growth and augmented cell width. Comparative RNA-seq analysis revealed that RnJ controls a large regulon encoding many factors predicted to be involved in biosynthesis, regulation, transport, and iron acquisition. One up-regulated gene in ∆rnj mutant is ftsH, coding for the cell division protein FtsH, an inner membrane protease. Interestingly, overexpression of FtsH in the wild-type strain also caused cell-width augmentation. However, unlike the rnj mutant, which was attenuated in a Caenorhabditis elegans model of infection, the FtsH-overexpressing strain exhibited the same virulence phenotype as the wild-type strain. Remarkably, under iron-depleted conditions, production of the exotoxin diphtheria toxin was significantly reduced in the rnj mutant compared to the wild-type strain, likely due to dysregulated secretion of the toxin. Evidently, RNase J is a key ribonuclease that post-transcriptionally influences the expression of factors vital to C. diphtheriae physiology and virulence. Overall design: Examination of RNase J deficient mutant compared to wildtype in biological triplicatesbacteriaENAAcid, Shape, viral infection, RNA, shape of cell, Ribonuclease, Microsporon diphtheriticum, Mycobacterium diphtheriae, Shapes, Exotoxin, infectivity, Cell, Microsporon diphthericum., RNase, Nucleases, Pacinia loeffleri, Bacillus diphtheriae, Alkaline Ribonuclease, Acid Ribonuclease, Pathogenicity, Bacterium diphtheriae, Cell Shapes, virulence, RNA Nucleases, AlkalineAcid, Shape, viral infection, RNA, shape of cell, Ribonuclease, Microsporon diphtheriticum, Mycobacterium diphtheriae, Shapes, Exotoxin, infectivity, Cell, Microsporon diphthericum., RNase, Nucleases, Pacinia loeffleri, Bacillus diphtheriae, Alkaline Ribonuclease, Acid Ribonuclease, Pathogenicity, Bacterium diphtheriae, Cell Shapes, virulence, RNA Nucleases, AlkalinefalseRibonuclease J modulates cell shape, exotoxin production, and virulence in Corynebacterium diphtheriaeRibonuclease J modulates cell shape, exotoxin production, and virulence in Corynebacterium diphtheriae2025-09-242021-02-11PRJNA694916GSE165533171733672886