Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43
Background: CpxAR is a two-component system that allows bacteria to reorganize envelope structures in response to extracellular stimuli. CpxAR negatively affects type 1 fimbriae expression in Klebsiella pneumoniae CG43, a hypervirulent strain. The involvement of CpxAR in the regulation of type 3 fim...
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| Format: | Article |
| Language: | English |
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Elsevier
2023-06-01
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| Series: | Journal of Microbiology, Immunology and Infection |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S1684118223000403 |
| _version_ | 1797812081455529984 |
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| author | Chih-Hao Kuo Wei-Feng Lin Chia-Jui Liu Zhe-Chong Wang Ting-Yi Liu Hwei-Ling Peng |
| author_facet | Chih-Hao Kuo Wei-Feng Lin Chia-Jui Liu Zhe-Chong Wang Ting-Yi Liu Hwei-Ling Peng |
| author_sort | Chih-Hao Kuo |
| collection | DOAJ |
| description | Background: CpxAR is a two-component system that allows bacteria to reorganize envelope structures in response to extracellular stimuli. CpxAR negatively affects type 1 fimbriae expression in Klebsiella pneumoniae CG43, a hypervirulent strain. The involvement of CpxAR in the regulation of type 3 fimbriae expression was investigated. Methods: cpxAR, cpxA, and cpxR gene-specific deletion mutants were generated. The deletion effects on the expression of type 1 and type 3 fimbriae were analyzed via measuring the promoter activity, mannose sensitive yeast agglutination activity, biofilm formation, and the production of the major pilins FimA and MrkA respectively. RNA sequencing analysis of CG43S3, ΔcpxAR, ΔcpxR and Δfur was employed to study the regulatory mechanism influencing the expression of type 3 fimbriae. Results: Deletion of cpxAR increased type 1 and type 3 fimbrial expression. Comparative transcriptomic analysis showed that the expression of oxidative stress-responsive enzymes, type 1 and type 3 fimbriae, and iron acquisition and homeostasis control systems were differentially affected by cpxAR or cpxR deletion. Subsequent analysis revealed that the small RNA RyhB negatively affects the expression of type 3 fimbriae, while CpxAR positively controls ryhB expression. Finally, the site-directed mutation of the predicted interacting sequences of RyhB with the mRNA of MrkA attenuated the RyhB repression of type 3 fimbriae. Conclusion: CpxAR negatively regulates the expression of type 3 fimbriae by modulating cellular iron levels thereafter activating the expression of RyhB. The activated RyhB represses the expression of type 3 fimbriae by base-pairing binding to the 5′region of mrkA mRNA. |
| first_indexed | 2024-03-13T07:32:17Z |
| format | Article |
| id | doaj.art-3935263bb5984093acb2ab14b48bd231 |
| institution | Directory Open Access Journal |
| issn | 1684-1182 |
| language | English |
| last_indexed | 2024-03-13T07:32:17Z |
| publishDate | 2023-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Journal of Microbiology, Immunology and Infection |
| spelling | doaj.art-3935263bb5984093acb2ab14b48bd2312023-06-04T04:23:27ZengElsevierJournal of Microbiology, Immunology and Infection1684-11822023-06-01563464476Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43Chih-Hao Kuo0Wei-Feng Lin1Chia-Jui Liu2Zhe-Chong Wang3Ting-Yi Liu4Hwei-Ling Peng5Department of Biological Science and Technology, School of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsin Chu, TaiwanInstitute of Molecular Medicine and Bioengineering, School of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsin Chu, TaiwanDepartment of Biological Science and Technology, School of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsin Chu, TaiwanDepartment of Biological Science and Technology, School of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsin Chu, TaiwanInstitute of Molecular Medicine and Bioengineering, School of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsin Chu, TaiwanDepartment of Biological Science and Technology, School of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsin Chu, Taiwan; Institute of Molecular Medicine and Bioengineering, School of Biological Science and Technology, National Yang Ming Chiao Tung University, Hsin Chu, Taiwan; Corresponding author. Department of Biological Science and Technology, National Yang Ming Chiao Tung University, 75 Po Ai Street, Hsin Chu, 30068, Taiwan.Background: CpxAR is a two-component system that allows bacteria to reorganize envelope structures in response to extracellular stimuli. CpxAR negatively affects type 1 fimbriae expression in Klebsiella pneumoniae CG43, a hypervirulent strain. The involvement of CpxAR in the regulation of type 3 fimbriae expression was investigated. Methods: cpxAR, cpxA, and cpxR gene-specific deletion mutants were generated. The deletion effects on the expression of type 1 and type 3 fimbriae were analyzed via measuring the promoter activity, mannose sensitive yeast agglutination activity, biofilm formation, and the production of the major pilins FimA and MrkA respectively. RNA sequencing analysis of CG43S3, ΔcpxAR, ΔcpxR and Δfur was employed to study the regulatory mechanism influencing the expression of type 3 fimbriae. Results: Deletion of cpxAR increased type 1 and type 3 fimbrial expression. Comparative transcriptomic analysis showed that the expression of oxidative stress-responsive enzymes, type 1 and type 3 fimbriae, and iron acquisition and homeostasis control systems were differentially affected by cpxAR or cpxR deletion. Subsequent analysis revealed that the small RNA RyhB negatively affects the expression of type 3 fimbriae, while CpxAR positively controls ryhB expression. Finally, the site-directed mutation of the predicted interacting sequences of RyhB with the mRNA of MrkA attenuated the RyhB repression of type 3 fimbriae. Conclusion: CpxAR negatively regulates the expression of type 3 fimbriae by modulating cellular iron levels thereafter activating the expression of RyhB. The activated RyhB represses the expression of type 3 fimbriae by base-pairing binding to the 5′region of mrkA mRNA.http://www.sciencedirect.com/science/article/pii/S1684118223000403CpxAR two-component systemIron homeostasis controlKlebsiella pneumoniae CG43Small RNA RyhBType 3 fimbriae expression |
| spellingShingle | Chih-Hao Kuo Wei-Feng Lin Chia-Jui Liu Zhe-Chong Wang Ting-Yi Liu Hwei-Ling Peng Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43 Journal of Microbiology, Immunology and Infection CpxAR two-component system Iron homeostasis control Klebsiella pneumoniae CG43 Small RNA RyhB Type 3 fimbriae expression |
| title | Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43 |
| title_full | Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43 |
| title_fullStr | Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43 |
| title_full_unstemmed | Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43 |
| title_short | Role of the stress-responsive two-component system CpxAR in regulating fimbriae expression in Klebsiella pneumoniae CG43 |
| title_sort | role of the stress responsive two component system cpxar in regulating fimbriae expression in klebsiella pneumoniae cg43 |
| topic | CpxAR two-component system Iron homeostasis control Klebsiella pneumoniae CG43 Small RNA RyhB Type 3 fimbriae expression |
| url | http://www.sciencedirect.com/science/article/pii/S1684118223000403 |
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