The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture.

Eilers, K; Hoong Yam, JK; Liu, X; Goh, YF; To, K-N; Paracuellos, P; Morton, R; Brizuela, J; Hui Yong, AM; Givskov, M; Freibert, S-A; Bange, G; Rice, SA; Steinchen, W; Filloux, A

The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture.

Eilers, K Hoong Yam, JK Liu, X Goh, YF To, K-N Paracuellos, P Morton, R Brizuela, J Hui Yong, AM Givskov, M Freibert, S-A Bange, G Rice, SA Steinchen, W Filloux, A

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Publisher:: ELSEVIER
Publication Type:: Journal Article
Citation:: J Biol Chem, 2024, 300, (2), pp. 105659
Issue Date:: 2024-02

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Field	Value	Language
dc.contributor.author	Eilers, K
dc.contributor.author	Hoong Yam, JK
dc.contributor.author	Liu, X
dc.contributor.author	Goh, YF
dc.contributor.author	To, K-N
dc.contributor.author	Paracuellos, P
dc.contributor.author	Morton, R
dc.contributor.author	Brizuela, J
dc.contributor.author	Hui Yong, AM
dc.contributor.author	Givskov, M
dc.contributor.author	Freibert, S-A
dc.contributor.author	Bange, G
dc.contributor.author	Rice, SA
dc.contributor.author	Steinchen, W
dc.contributor.author	Filloux, A
dc.date.accessioned	2024-12-02T00:38:19Z
dc.date.available	2024-01-04
dc.date.available	2024-12-02T00:38:19Z
dc.date.issued	2024-02
dc.identifier.citation	J Biol Chem, 2024, 300, (2), pp. 105659
dc.identifier.issn	0021-9258
dc.identifier.issn	1083-351X
dc.identifier.uri	http://hdl.handle.net/10453/182159
dc.description.abstract	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.
dc.format	Print-Electronic
dc.language	eng
dc.publisher	ELSEVIER
dc.relation.ispartof	J Biol Chem
dc.relation.isbasedon	10.1016/j.jbc.2024.105659
dc.rights	info:eu-repo/semantics/openAccess
dc.subject	03 Chemical Sciences, 06 Biological Sciences, 11 Medical and Health Sciences
dc.subject.classification	Biochemistry & Molecular Biology
dc.subject.classification	31 Biological sciences
dc.subject.classification	32 Biomedical and clinical sciences
dc.subject.classification	34 Chemical sciences
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Biofilms
dc.subject.mesh	Cyclic GMP
dc.subject.mesh	Gene Expression Regulation, Bacterial
dc.subject.mesh	Phosphoric Diester Hydrolases
dc.subject.mesh	Phosphorus-Oxygen Lyases
dc.subject.mesh	Pseudomonas
dc.subject.mesh	Biofilms
dc.subject.mesh	Pseudomonas
dc.subject.mesh	Phosphoric Diester Hydrolases
dc.subject.mesh	Phosphorus-Oxygen Lyases
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Cyclic GMP
dc.subject.mesh	Gene Expression Regulation, Bacterial
dc.subject.mesh	Bacterial Proteins
dc.subject.mesh	Biofilms
dc.subject.mesh	Cyclic GMP
dc.subject.mesh	Gene Expression Regulation, Bacterial
dc.subject.mesh	Phosphoric Diester Hydrolases
dc.subject.mesh	Phosphorus-Oxygen Lyases
dc.subject.mesh	Pseudomonas
dc.title	The dual GGDEF/EAL domain enzyme PA0285 is a Pseudomonas species housekeeping phosphodiesterase regulating early attachment and biofilm architecture.
dc.type	Journal Article
utslib.citation.volume	300
utslib.location.activity	United States
utslib.for	03 Chemical Sciences
utslib.for	06 Biological Sciences
utslib.for	11 Medical and Health Sciences
pubs.organisational-group	University of Technology Sydney
pubs.organisational-group	University of Technology Sydney/Faculty of Science
pubs.organisational-group	University of Technology Sydney/UTS Groups
pubs.organisational-group	University of Technology Sydney/UTS Groups/Australian Institute for Microbiology & Infection (AIMI)
pubs.organisational-group	University of Technology Sydney/UTS Groups/Australian Institute for Microbiology & Infection (AIMI)/Australian Institute for Microbiology & Infection (AIMI) Associate Members
utslib.copyright.status	open_access	*
dc.rights.license	This work is licensed under a Creative Commons Attribution 4.0 International License (CC BY 4.0). To view a copy of this license, visit https://creativecommons.org/licenses/by/4.0/
dc.date.updated	2024-12-02T00:38:13Z
pubs.issue	2
pubs.publication-status	Published
pubs.volume	300
utslib.citation.issue	2

Abstract:

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.

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http://hdl.handle.net/10453/182159