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| Status |
Public on Jul 17, 2024 |
| Title |
RNA interactome profiling in hypervirulent Klebsiella pneumoniae reveals small RNA ArcZ as an inhibitor of mucoviscosity and virulence |
| Organism |
Klebsiella pneumoniae subsp. pneumoniae ATCC 43816 |
| Experiment type |
Expression profiling by high throughput sequencing
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| Summary |
Hypervirulent Klebsiella pneumoniae (HvKP) is an emerging human pathogen causing invasive infection in immune-competent hosts. The hypervirulence is strongly linked to the overproduction of hypermucovisous capsule, but the underlining regulatory mechanism of hypermucoviscosity (HMV) has been elusive, especially at the post-transcriptional level mediated by small RNAs (sRNAs). Using a recently developed RNA interactome profiling approach, we have investigated the Hfq-associated sRNA regulatory network and established the first in vivo RNA-RNA interactome in HvKP. Our data reveal numerous interactions between sRNAs and HMV-related mRNAs, and identify a plethora of sRNA that inhibit or promote HMV. One of the strongest repressors of HMV was ArcZ, a conserved sRNA in the Enterobacteriaceae family. We found that ArcZ is activated by the master regulator of catabolite repression Crp, and down-regulates the expression of mlaA encoding an outer-membrane lipoprotein, leading to decreased HMV and virulence attenuation in mice. ArcZ significantly reduced HMV in several carbapenem-resistant and hypervirulent clinical isolates with diverse genetic background, suggesting it is an antisense RNA inhibitor of HMV with therapeutic potential. In summary, our work provides a comprehensive map of the RNA-RNA interaction network of HvKP and identifies ArcZ as a conserved repressor of HMV, providing novel insights into the mechanisms of posttranscriptional regulations of virulence.
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| Overall design |
To assess the the effect of expression of ArcZ and MlaA on transcriptome, we have first applied RNA-seq to biological triplicates of K. pneumoniae WT, ΔmlaA, ΔarcZ, ΔarcZ+pZE12-ArcZ and ΔarcZ+pZE12-ArcZ-M5. WT and ΔarcZ carry the empty vector. Strains were grown in LB media containing IPTG as sRNA expression inducer. Cells at OD600 3 were harvested to construct the strand-specific cDNA library and to be sequenced.
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| Contributor(s) |
Chao Y, Wu K |
| Citation(s) |
39138169 |
| |
| Submission date |
Mar 03, 2024 |
| Last update date |
Sep 12, 2024 |
| Contact name |
Kejing Wu |
| E-mail(s) |
wukejing104@126.com
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| Organization name |
Shanghai Institute of Immunity and Infection, Chinese Academy of Sciences
|
| Street address |
320 Yueyang Road
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| City |
Shanghai |
| ZIP/Postal code |
200031 |
| Country |
China |
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| Platforms (1) |
| GPL33764 |
Illumina NovaSeq 6000 (Klebsiella pneumoniae subsp. pneumoniae ATCC 43816) |
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| Samples (14)
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| Relations |
| BioProject |
PRJNA1083197 |
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