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RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa

Pseudomonas aeruginosa is an important opportunistic pathogen capable of causing variety of infections in humans. The type III secretion system (T3SS) is a critical virulence determinant of P. aeruginosa in the host infections. Expression of the T3SS is regulated by ExsA, a master regulator that act...

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Autores principales: Wang, Dan, Zhang, Xinxin, Yin, Liwen, Liu, Qi, Yu, Zhaoli, Xu, Congjuan, Ma, Zhenzhen, Xia, Yushan, Shi, Jing, Gong, Yuehua, Bai, Fang, Cheng, Zhihui, Wu, Weihui, Lin, Jinzhong, Jin, Yongxin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8730436/
https://www.ncbi.nlm.nih.gov/pubmed/34986198
http://dx.doi.org/10.1371/journal.ppat.1010170
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author Wang, Dan
Zhang, Xinxin
Yin, Liwen
Liu, Qi
Yu, Zhaoli
Xu, Congjuan
Ma, Zhenzhen
Xia, Yushan
Shi, Jing
Gong, Yuehua
Bai, Fang
Cheng, Zhihui
Wu, Weihui
Lin, Jinzhong
Jin, Yongxin
author_facet Wang, Dan
Zhang, Xinxin
Yin, Liwen
Liu, Qi
Yu, Zhaoli
Xu, Congjuan
Ma, Zhenzhen
Xia, Yushan
Shi, Jing
Gong, Yuehua
Bai, Fang
Cheng, Zhihui
Wu, Weihui
Lin, Jinzhong
Jin, Yongxin
author_sort Wang, Dan
collection PubMed
description Pseudomonas aeruginosa is an important opportunistic pathogen capable of causing variety of infections in humans. The type III secretion system (T3SS) is a critical virulence determinant of P. aeruginosa in the host infections. Expression of the T3SS is regulated by ExsA, a master regulator that activates the expression of all known T3SS genes. Expression of the exsA gene is controlled at both transcriptional and posttranscriptional levels. Here, we screened a P. aeruginosa transposon (Tn5) insertional mutant library and found rplI, a gene coding for the ribosomal large subunit protein L9, to be a repressor for the T3SS gene expression. Combining real-time quantitative PCR (qPCR), western blotting and lacZ fusion assays, we show that RplI controls the expression of exsA at the posttranscriptional level. Further genetic experiments demonstrated that RplI mediated control of the exsA translation involves 5’ untranslated region (5’ UTR). A ribosome immunoprecipitation assay and qPCR revealed higher amounts of a 24 nt fragment from exsA mRNA being associated with ribosomes in the ΔrplI mutant. An interaction between RplI and exsA mRNA harboring its 24 nt, but not 12 nt, 5’ UTR was confirmed by RNA Gel Mobility Shift and Microscale Thermophoresis assays. Overall, this study identifies the ribosomal large subunit protein L9 as a novel T3SS repressor that inhibits ExsA translation in P. aeruginosa.
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spelling pubmed-87304362022-01-06 RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa Wang, Dan Zhang, Xinxin Yin, Liwen Liu, Qi Yu, Zhaoli Xu, Congjuan Ma, Zhenzhen Xia, Yushan Shi, Jing Gong, Yuehua Bai, Fang Cheng, Zhihui Wu, Weihui Lin, Jinzhong Jin, Yongxin PLoS Pathog Research Article Pseudomonas aeruginosa is an important opportunistic pathogen capable of causing variety of infections in humans. The type III secretion system (T3SS) is a critical virulence determinant of P. aeruginosa in the host infections. Expression of the T3SS is regulated by ExsA, a master regulator that activates the expression of all known T3SS genes. Expression of the exsA gene is controlled at both transcriptional and posttranscriptional levels. Here, we screened a P. aeruginosa transposon (Tn5) insertional mutant library and found rplI, a gene coding for the ribosomal large subunit protein L9, to be a repressor for the T3SS gene expression. Combining real-time quantitative PCR (qPCR), western blotting and lacZ fusion assays, we show that RplI controls the expression of exsA at the posttranscriptional level. Further genetic experiments demonstrated that RplI mediated control of the exsA translation involves 5’ untranslated region (5’ UTR). A ribosome immunoprecipitation assay and qPCR revealed higher amounts of a 24 nt fragment from exsA mRNA being associated with ribosomes in the ΔrplI mutant. An interaction between RplI and exsA mRNA harboring its 24 nt, but not 12 nt, 5’ UTR was confirmed by RNA Gel Mobility Shift and Microscale Thermophoresis assays. Overall, this study identifies the ribosomal large subunit protein L9 as a novel T3SS repressor that inhibits ExsA translation in P. aeruginosa. Public Library of Science 2022-01-05 /pmc/articles/PMC8730436/ /pubmed/34986198 http://dx.doi.org/10.1371/journal.ppat.1010170 Text en © 2022 Wang et al https://creativecommons.org/licenses/by/4.0/This is an open access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Wang, Dan
Zhang, Xinxin
Yin, Liwen
Liu, Qi
Yu, Zhaoli
Xu, Congjuan
Ma, Zhenzhen
Xia, Yushan
Shi, Jing
Gong, Yuehua
Bai, Fang
Cheng, Zhihui
Wu, Weihui
Lin, Jinzhong
Jin, Yongxin
RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa
title RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa
title_full RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa
title_fullStr RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa
title_full_unstemmed RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa
title_short RplI interacts with 5’ UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa
title_sort rpli interacts with 5’ utr of exsa to repress its translation and type iii secretion system in pseudomonas aeruginosa
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8730436/
https://www.ncbi.nlm.nih.gov/pubmed/34986198
http://dx.doi.org/10.1371/journal.ppat.1010170
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