The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture
Bacterial lifestyles depend on conditions encountered during colonization. The transition between planktonic and biofilm growth is dependent on the intracellular second messenger c-di-GMP. High c-di-GMP levels driven by diguanylate cyclases (DGCs) activity favor biofilm formation, while low levels w...
| Main Authors: | , , , , , , , , , , , , , , |
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| Other Authors: | |
| Format: | Journal Article |
| Language: | English |
| Published: |
2024
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| Subjects: | |
| Online Access: | https://hdl.handle.net/10356/178529 |
| _version_ | 1824456148402569216 |
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| author | Eilers, Kira Yam, Joey Kuok Hoong Liu, Xianghui Goh, Yu Fen To, Ka-Ning Paracuellos, Patricia Morton, Richard Brizuela, Jaime Yong, Adeline Mei Hui Givskov, Michael Freibert, Sven-Andreas Bange, Gert Rice, Scott A. Steinchen, Wieland Filloux, Alain |
| author2 | Singapore Centre for Environmental Life Sciences and Engineering |
| author_facet | Singapore Centre for Environmental Life Sciences and Engineering Eilers, Kira Yam, Joey Kuok Hoong Liu, Xianghui Goh, Yu Fen To, Ka-Ning Paracuellos, Patricia Morton, Richard Brizuela, Jaime Yong, Adeline Mei Hui Givskov, Michael Freibert, Sven-Andreas Bange, Gert Rice, Scott A. Steinchen, Wieland Filloux, Alain |
| author_sort | Eilers, Kira |
| collection | NTU |
| description | Bacterial lifestyles depend on conditions encountered during colonization. The transition between planktonic and biofilm growth is dependent on the intracellular second messenger c-di-GMP. High c-di-GMP levels driven by diguanylate cyclases (DGCs) activity favor biofilm formation, while low levels were maintained by phosphodiesterases (PDE) encourage planktonic lifestyle. The activity of these enzymes can be modulated by stimuli-sensing domains such as Per-ARNT-Sim (PAS). In Pseudomonas aeruginosa, more than 40 PDE/DGC are involved in c-di-GMP homeostasis, including 16 dual proteins possessing both canonical DGC and PDE motifs, that is, GGDEF and EAL, respectively. It was reported that deletion of the EAL/GGDEF dual enzyme PA0285, one of five c-di-GMP-related enzymes conserved across all Pseudomonas species, impacts biofilms. PA0285 is anchored in the membrane and carries two PAS domains. Here, we confirm that its role is conserved in various P. aeruginosa strains and in Pseudomonas putida. Deletion of PA0285 impacts the early stage of colonization, and RNA-seq analysis suggests that expression of cupA fimbrial genes is involved. We demonstrate that the C-terminal portion of PA0285 encompassing the GGDEF and EAL domains binds GTP and c-di-GMP, respectively, but only exhibits PDE activity in vitro. However, both GGDEF and EAL domains are important for PA0285 PDE activity in vivo. Complementation of the PA0285 mutant strain with a copy of the gene encoding the C-terminal GGDEF/EAL portion in trans was not as effective as complementation with the full-length gene. This suggests the N-terminal transmembrane and PAS domains influence the PDE activity in vivo, through modulating the protein conformation. |
| first_indexed | 2025-02-19T03:49:30Z |
| format | Journal Article |
| id | ntu-10356/178529 |
| institution | Nanyang Technological University |
| language | English |
| last_indexed | 2025-02-19T03:49:30Z |
| publishDate | 2024 |
| record_format | dspace |
| spelling | ntu-10356/1785292024-06-27T15:30:26Z The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture Eilers, Kira Yam, Joey Kuok Hoong Liu, Xianghui Goh, Yu Fen To, Ka-Ning Paracuellos, Patricia Morton, Richard Brizuela, Jaime Yong, Adeline Mei Hui Givskov, Michael Freibert, Sven-Andreas Bange, Gert Rice, Scott A. Steinchen, Wieland Filloux, Alain Singapore Centre for Environmental Life Sciences and Engineering Medicine, Health and Life Sciences Diguanylate cyclase Phosphodiesterase Bacterial lifestyles depend on conditions encountered during colonization. The transition between planktonic and biofilm growth is dependent on the intracellular second messenger c-di-GMP. High c-di-GMP levels driven by diguanylate cyclases (DGCs) activity favor biofilm formation, while low levels were maintained by phosphodiesterases (PDE) encourage planktonic lifestyle. The activity of these enzymes can be modulated by stimuli-sensing domains such as Per-ARNT-Sim (PAS). In Pseudomonas aeruginosa, more than 40 PDE/DGC are involved in c-di-GMP homeostasis, including 16 dual proteins possessing both canonical DGC and PDE motifs, that is, GGDEF and EAL, respectively. It was reported that deletion of the EAL/GGDEF dual enzyme PA0285, one of five c-di-GMP-related enzymes conserved across all Pseudomonas species, impacts biofilms. PA0285 is anchored in the membrane and carries two PAS domains. Here, we confirm that its role is conserved in various P. aeruginosa strains and in Pseudomonas putida. Deletion of PA0285 impacts the early stage of colonization, and RNA-seq analysis suggests that expression of cupA fimbrial genes is involved. We demonstrate that the C-terminal portion of PA0285 encompassing the GGDEF and EAL domains binds GTP and c-di-GMP, respectively, but only exhibits PDE activity in vitro. However, both GGDEF and EAL domains are important for PA0285 PDE activity in vivo. Complementation of the PA0285 mutant strain with a copy of the gene encoding the C-terminal GGDEF/EAL portion in trans was not as effective as complementation with the full-length gene. This suggests the N-terminal transmembrane and PAS domains influence the PDE activity in vivo, through modulating the protein conformation. Ministry of Education (MOE) National Research Foundation (NRF) Published version The authors would like to acknowledge the financial support from National Research Foundation and Ministry of Education Singapore under its Research Centre of Excellence Program. A. F. work was supported by BBSRC grant BB/R00174X/1. G. B. received support by the Deutsche Forschungsgemeinschaft (DFG) for the core facility for Interactions, Dynamics, and Macromolecular Assembly (project 324652314). R. M. was supported by a MRC Clinical Research Training Fellowship. 2024-06-25T05:21:16Z 2024-06-25T05:21:16Z 2024 Journal Article Eilers, K., Yam, J. K. H., Liu, X., Goh, Y. F., To, K., Paracuellos, P., Morton, R., Brizuela, J., Yong, A. M. H., Givskov, M., Freibert, S., Bange, G., Rice, S. A., Steinchen, W. & Filloux, A. (2024). The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture. Journal of Biological Chemistry, 300(2), 105659-. https://dx.doi.org/10.1016/j.jbc.2024.105659 0021-9258 https://hdl.handle.net/10356/178529 10.1016/j.jbc.2024.105659 38237678 2-s2.0-85184665766 2 300 105659 en Journal of Biological Chemistry © 2024 The Authors. Published by Elsevier Inc on behalf of American Society for Biochemistry and Molecular Biology. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). application/pdf |
| spellingShingle | Medicine, Health and Life Sciences Diguanylate cyclase Phosphodiesterase Eilers, Kira Yam, Joey Kuok Hoong Liu, Xianghui Goh, Yu Fen To, Ka-Ning Paracuellos, Patricia Morton, Richard Brizuela, Jaime Yong, Adeline Mei Hui Givskov, Michael Freibert, Sven-Andreas Bange, Gert Rice, Scott A. Steinchen, Wieland Filloux, Alain The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture |
| title | The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture |
| title_full | The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture |
| title_fullStr | The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture |
| title_full_unstemmed | The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture |
| title_short | The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture |
| title_sort | dual ggdef eal domain enzyme pa0285 is a pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture |
| topic | Medicine, Health and Life Sciences Diguanylate cyclase Phosphodiesterase |
| url | https://hdl.handle.net/10356/178529 |
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