Insights into biofilm dispersal regulation from the crystal structure of the PAS-GGDEF-EAL region of RbdA from Pseudomonas aeruginosa

Liu, C; Liew, CW; Wong, YH; Tan, ST; Poh, WH; Manimekalai, MSS; Rajan, S; Xin, L; Liang, ZX; Grüber, G; Rice, SA; Lescar, J

Insights into biofilm dispersal regulation from the crystal structure of the PAS-GGDEF-EAL region of RbdA from Pseudomonas aeruginosa

Liu, C Liew, CW Wong, YH Tan, ST Poh, WH Manimekalai, MSS Rajan, S Xin, L Liang, ZX Grüber, G Rice, SA

Lescar, J

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Publication Type:: Journal Article
Citation:: Journal of Bacteriology, 2018, 200 (3)
Issue Date:: 2018-02-01

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Field	Value	Language
dc.contributor.author	Liu, C	en_US
dc.contributor.author	Liew, CW	en_US
dc.contributor.author	Wong, YH	en_US
dc.contributor.author	Tan, ST	en_US
dc.contributor.author	Poh, WH	en_US
dc.contributor.author	Manimekalai, MSS	en_US
dc.contributor.author	Rajan, S	en_US
dc.contributor.author	Xin, L	en_US
dc.contributor.author	Liang, ZX	en_US
dc.contributor.author	Grüber, G	en_US
dc.contributor.author	Rice, SA https://orcid.org/0000-0002-9486-2343	en_US
dc.contributor.author	Lescar, J	en_US
dc.date.available	2017-10-26	en_US
dc.date.issued	2018-02-01	en_US
dc.identifier.citation	Journal of Bacteriology, 2018, 200 (3)	en_US
dc.identifier.issn	0021-9193	en_US
dc.identifier.uri	http://hdl.handle.net/10453/122476
dc.description.abstract	© 2018 American Society for Microbiology. RbdA is a positive regulator of biofilm dispersal of Pseudomonas aeruginosa. Its cytoplasmic region (cRbdA) comprises an N-terminal Per-ARNT-Sim (PAS) domain followed by a diguanylate cyclase (GGDEF) domain and an EAL domain, whose phosphodiesterase activity is allosterically stimulated by GTP binding to the GGDEF domain. We report crystal structures of cRbdA and of two binary complexes: one with GTP/Mg 2+ bound to the GGDEF active site and one with the EAL domain bound to the c-di-GMP substrate. These structures unveil a 2-fold symmetric dimer stabilized by a closely packed N-terminal PAS domain and a noncanonical EAL dimer. The autoinhibitory switch is formed by an α-helix (S-helix) immediately N-terminal to the GGDEF domain that interacts with the EAL dimerization helix (α6-E) of the other EAL monomer and maintains the protein in a locked conformation. We propose that local conformational changes in cRbdA upon GTP binding lead to a structure with the PAS domain and S-helix shifted away from the GGDEF-EAL domains, as suggested by small-angle X-ray scattering (SAXS) experiments. Domain reorientation should be facilitated by the presence of an α-helical lever (H-helix) that tethers the GGDEF and EAL regions, allowing the EAL domain to rearrange into an active dimeric conformation.	en_US
dc.relation.ispartof	Journal of Bacteriology	en_US
dc.relation.isbasedon	10.1128/JB.00515-17	en_US
dc.subject.classification	Microbiology	en_US
dc.subject.mesh	Biofilms	en_US
dc.subject.mesh	Pseudomonas aeruginosa	en_US
dc.subject.mesh	Bacterial Proteins	en_US
dc.subject.mesh	Crystallography	en_US
dc.subject.mesh	Cloning, Molecular	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Protein Conformation	en_US
dc.subject.mesh	Models, Molecular	en_US
dc.subject.mesh	Protein Domains	en_US
dc.title	Insights into biofilm dispersal regulation from the crystal structure of the PAS-GGDEF-EAL region of RbdA from Pseudomonas aeruginosa	en_US
dc.type	Journal Article
utslib.citation.volume	3	en_US
utslib.citation.volume	200	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	1101 Medical Biochemistry and Metabolomics	en_US
utslib.for	06 Biological Sciences	en_US
utslib.for	07 Agricultural and Veterinary Sciences	en_US
utslib.for	11 Medical and Health Sciences	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Strength - ithree - Institute of Infection, Immunity and Innovation
utslib.copyright.status	open_access
pubs.issue	3	en_US
pubs.publication-status	Published	en_US
pubs.volume	200	en_US

Abstract:

© 2018 American Society for Microbiology. RbdA is a positive regulator of biofilm dispersal of Pseudomonas aeruginosa. Its cytoplasmic region (cRbdA) comprises an N-terminal Per-ARNT-Sim (PAS) domain followed by a diguanylate cyclase (GGDEF) domain and an EAL domain, whose phosphodiesterase activity is allosterically stimulated by GTP binding to the GGDEF domain. We report crystal structures of cRbdA and of two binary complexes: one with GTP/Mg 2+ bound to the GGDEF active site and one with the EAL domain bound to the c-di-GMP substrate. These structures unveil a 2-fold symmetric dimer stabilized by a closely packed N-terminal PAS domain and a noncanonical EAL dimer. The autoinhibitory switch is formed by an α-helix (S-helix) immediately N-terminal to the GGDEF domain that interacts with the EAL dimerization helix (α6-E) of the other EAL monomer and maintains the protein in a locked conformation. We propose that local conformational changes in cRbdA upon GTP binding lead to a structure with the PAS domain and S-helix shifted away from the GGDEF-EAL domains, as suggested by small-angle X-ray scattering (SAXS) experiments. Domain reorientation should be facilitated by the presence of an α-helical lever (H-helix) that tethers the GGDEF and EAL regions, allowing the EAL domain to rearrange into an active dimeric conformation.

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http://hdl.handle.net/10453/122476