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| Status |
Public on Feb 08, 2021 |
| Title |
Ribonuclease J modulates cell shape, exotoxin production, and virulence in Corynebacterium diphtheriae |
| Organism |
Corynebacterium diphtheriae |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
RNA 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.
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| Overall design |
Examination of RNase J deficient mutant compared to wildtype in biological triplicates
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| Contributor(s) |
Luong TT, Nguyen MT, Chen Y, Chang C, Lee JH, Wittchen M, Ton-That H, Cruz M, Garsin DA, Das A, Tauch A, Ton-That H |
| Citation(s) |
33672886 |
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| Submission date |
Jan 26, 2021 |
| Last update date |
Mar 11, 2021 |
| Contact name |
Manuel Wittchen |
| Organization name |
Bielefeld University
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| Department |
Center for Biotechnology
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| Street address |
Universitaetsstr. 27
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| City |
Bielefeld |
| ZIP/Postal code |
33615 |
| Country |
Germany |
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| Platforms (1) |
| GPL29654 |
Illumina MiSeq (Corynebacterium diphtheriae) |
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| Samples (6)
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| Relations |
| BioProject |
PRJNA694916 |
| SRA |
SRP303319 |
| Supplementary file |
Size |
Download |
File type/resource |
| GSE165533_List_of_differentially_transcribed_genes.xlsx |
340.1 Kb |
(ftp)(http) |
XLSX |
SRA Run Selector |
| Raw data are available in SRA |
| Processed data are available on Series record |
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