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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Public Library of Science (PLoS)
2022-01-01
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| Series: | PLoS Pathogens |
| Online Access: | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010170&type=printable |
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| author | Dan Wang Xinxin Zhang Liwen Yin Qi Liu Zhaoli Yu Congjuan Xu Zhenzhen Ma Yushan Xia Jing Shi Yuehua Gong Fang Bai Zhihui Cheng Weihui Wu Jinzhong Lin Yongxin Jin |
| author_facet | Dan Wang Xinxin Zhang Liwen Yin Qi Liu Zhaoli Yu Congjuan Xu Zhenzhen Ma Yushan Xia Jing Shi Yuehua Gong Fang Bai Zhihui Cheng Weihui Wu Jinzhong Lin Yongxin Jin |
| author_sort | Dan Wang |
| collection | DOAJ |
| 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. |
| format | Article |
| id | doaj-art-ff7ff6b41ee84a538edd09d63f56f4db |
| institution | DOAJ |
| issn | 1553-7366 1553-7374 |
| language | English |
| publishDate | 2022-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS Pathogens |
| spelling | doaj-art-ff7ff6b41ee84a538edd09d63f56f4db2025-08-20T03:16:25ZengPublic Library of Science (PLoS)PLoS Pathogens1553-73661553-73742022-01-01181e101017010.1371/journal.ppat.1010170RplI interacts with 5' UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa.Dan WangXinxin ZhangLiwen YinQi LiuZhaoli YuCongjuan XuZhenzhen MaYushan XiaJing ShiYuehua GongFang BaiZhihui ChengWeihui WuJinzhong LinYongxin JinPseudomonas 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.https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010170&type=printable |
| spellingShingle | Dan Wang Xinxin Zhang Liwen Yin Qi Liu Zhaoli Yu Congjuan Xu Zhenzhen Ma Yushan Xia Jing Shi Yuehua Gong Fang Bai Zhihui Cheng Weihui Wu Jinzhong Lin Yongxin Jin RplI interacts with 5' UTR of exsA to repress its translation and type III secretion system in Pseudomonas aeruginosa. PLoS Pathogens |
| 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 |
| url | https://journals.plos.org/plospathogens/article/file?id=10.1371/journal.ppat.1010170&type=printable |
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