Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1
ABSTRACT Several key virulence factors of Pseudomonas aeruginosa are regulated by quorum-sensing systems, small noncoding RNAs (sRNAs), and environmental stress, leading to a high mortality rate. Our previous studies indicated that the alkaline phosphatase LapA regulated P. aeruginosa PAO1 biofilm f...
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American Society for Microbiology
2025-06-01
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| Series: | Microbiology Spectrum |
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| Online Access: | https://journals.asm.org/doi/10.1128/spectrum.01303-24 |
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| author | Xiaojuan Tan Jingjing Xiao Qianqian Liu Ting Yang Dandan Feng Ruyi Zheng Liping Luo Xi Cheng Dongsheng Du Minghui Li Jinwei Zhou Guoping Zhu |
| author_facet | Xiaojuan Tan Jingjing Xiao Qianqian Liu Ting Yang Dandan Feng Ruyi Zheng Liping Luo Xi Cheng Dongsheng Du Minghui Li Jinwei Zhou Guoping Zhu |
| author_sort | Xiaojuan Tan |
| collection | DOAJ |
| description | ABSTRACT Several key virulence factors of Pseudomonas aeruginosa are regulated by quorum-sensing systems, small noncoding RNAs (sRNAs), and environmental stress, leading to a high mortality rate. Our previous studies indicated that the alkaline phosphatase LapA regulated P. aeruginosa PAO1 biofilm formation in a chronic wound model established with ex vivo porcine skin explants. Notably, one particular sRNA located upstream of the lapA gene was highly expressed in the model. Therefore, the sRNA was further characterized via northern blotting and rapid amplification of cDNA ends. The results revealed that the sRNA we named LapS is 197 nucleotides in length and is derived from the 5´ UTR and sequence internal to the lapA gene. Next, LapS mutation, overexpression, and complementation strains were constructed from the PAO1 strain, and phenotypic experiments associated with lapA were performed and compared with those of the ΔlapA and wild-type strains. The results indicated that LapS is involved in regulating swarming motility, rhamnolipid and alkaline phosphatase production, las/rhl quorum-sensing systems, and biofilm formation by controlling the level of lapA mRNA under phosphate-depleted conditions. Therefore, this LapS-lapA signaling cascade is beneficial for balancing the virulence regulation of P. aeruginosa. Additionally, an in vitro study indicated that LapS directly and post-transcriptionally regulated at least one unlinked gene, putA, which encodes bifunctional proline dehydrogenase, a virulence factor of P. aeruginosa. Collectively, our findings reveal a previously unstudied regulatory sRNA and advance the understanding of the roles of sRNAs in the pathogenicity of P. aeruginosa.IMPORTANCEPseudomonas aeruginosa is a common nosocomial pathogen that contains hundreds of virulence factors regulated by quorum-sensing systems and environmental stress. Small noncoding RNAs (sRNAs) involved in virulence regulation have been identified in P. aeruginosa. Recently, several potential sRNAs were identified in P. aeruginosa using transcriptome sequencing. However, some of these novel sRNAs have been functionally characterized. In this study, a previously uncharacterized sRNA, LapS, in P. aeruginosa PAO1 was identified as a novel sRNA. LapS is involved in regulating swarming motility, rhamnolipid production, and alkaline phosphatase production during phosphate-depleted stress by controlling the level of lapA mRNA. Furthermore, LapS deletion also reduced the mortality rate of Caenorhabditis elegans in a fast-kill assay. Additionally, LapS directly suppressed PutA, a virulence factor of P. aeruginosa. This study highlights the role of LapS in modulating P. aeruginosa virulence during phosphate-depleted stress. |
| format | Article |
| id | doaj-art-91534d7cecdf48e8a4cacd7ece5837ca |
| institution | OA Journals |
| issn | 2165-0497 |
| language | English |
| publishDate | 2025-06-01 |
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| spelling | doaj-art-91534d7cecdf48e8a4cacd7ece5837ca2025-08-20T02:05:24ZengAmerican Society for MicrobiologyMicrobiology Spectrum2165-04972025-06-0113610.1128/spectrum.01303-24Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1Xiaojuan Tan0Jingjing Xiao1Qianqian Liu2Ting Yang3Dandan Feng4Ruyi Zheng5Liping Luo6Xi Cheng7Dongsheng Du8Minghui Li9Jinwei Zhou10Guoping Zhu11Anhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaSchool of Food and Biological Engineering, Xuzhou University of Technology, Xuzhou, Jiangsu, ChinaAnhui Provincial Key Laboratory of Molecular Enzymology and Mechanism of Major Diseases, College of Life Sciences, Anhui Normal University, Wuhu, Anhui, ChinaABSTRACT Several key virulence factors of Pseudomonas aeruginosa are regulated by quorum-sensing systems, small noncoding RNAs (sRNAs), and environmental stress, leading to a high mortality rate. Our previous studies indicated that the alkaline phosphatase LapA regulated P. aeruginosa PAO1 biofilm formation in a chronic wound model established with ex vivo porcine skin explants. Notably, one particular sRNA located upstream of the lapA gene was highly expressed in the model. Therefore, the sRNA was further characterized via northern blotting and rapid amplification of cDNA ends. The results revealed that the sRNA we named LapS is 197 nucleotides in length and is derived from the 5´ UTR and sequence internal to the lapA gene. Next, LapS mutation, overexpression, and complementation strains were constructed from the PAO1 strain, and phenotypic experiments associated with lapA were performed and compared with those of the ΔlapA and wild-type strains. The results indicated that LapS is involved in regulating swarming motility, rhamnolipid and alkaline phosphatase production, las/rhl quorum-sensing systems, and biofilm formation by controlling the level of lapA mRNA under phosphate-depleted conditions. Therefore, this LapS-lapA signaling cascade is beneficial for balancing the virulence regulation of P. aeruginosa. Additionally, an in vitro study indicated that LapS directly and post-transcriptionally regulated at least one unlinked gene, putA, which encodes bifunctional proline dehydrogenase, a virulence factor of P. aeruginosa. Collectively, our findings reveal a previously unstudied regulatory sRNA and advance the understanding of the roles of sRNAs in the pathogenicity of P. aeruginosa.IMPORTANCEPseudomonas aeruginosa is a common nosocomial pathogen that contains hundreds of virulence factors regulated by quorum-sensing systems and environmental stress. Small noncoding RNAs (sRNAs) involved in virulence regulation have been identified in P. aeruginosa. Recently, several potential sRNAs were identified in P. aeruginosa using transcriptome sequencing. However, some of these novel sRNAs have been functionally characterized. In this study, a previously uncharacterized sRNA, LapS, in P. aeruginosa PAO1 was identified as a novel sRNA. LapS is involved in regulating swarming motility, rhamnolipid production, and alkaline phosphatase production during phosphate-depleted stress by controlling the level of lapA mRNA. Furthermore, LapS deletion also reduced the mortality rate of Caenorhabditis elegans in a fast-kill assay. Additionally, LapS directly suppressed PutA, a virulence factor of P. aeruginosa. This study highlights the role of LapS in modulating P. aeruginosa virulence during phosphate-depleted stress.https://journals.asm.org/doi/10.1128/spectrum.01303-24Pseudomonas aeruginosasRNA LapSalkaline phosphatase LapAvirulence factorsbiofilms |
| spellingShingle | Xiaojuan Tan Jingjing Xiao Qianqian Liu Ting Yang Dandan Feng Ruyi Zheng Liping Luo Xi Cheng Dongsheng Du Minghui Li Jinwei Zhou Guoping Zhu Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1 Microbiology Spectrum Pseudomonas aeruginosa sRNA LapS alkaline phosphatase LapA virulence factors biofilms |
| title | Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1 |
| title_full | Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1 |
| title_fullStr | Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1 |
| title_full_unstemmed | Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1 |
| title_short | Regulatory roles of an sRNA derived from the 5´ UTR and sequence internal to lapA in Pseudomonas aeruginosa PAO1 |
| title_sort | regulatory roles of an srna derived from the 5´ utr and sequence internal to lapa in pseudomonas aeruginosa pao1 |
| topic | Pseudomonas aeruginosa sRNA LapS alkaline phosphatase LapA virulence factors biofilms |
| url | https://journals.asm.org/doi/10.1128/spectrum.01303-24 |
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