title: Dcsbis (PA2771) from Pseudomonas aeruginosa is a highly active diguanylate cyclase with unique activity regulation.
abstract: C-di-GMP (3',5' -Cyclic diguanylic acid) is an important second messenger in bacteria that influences virulence, motility, biofilm formation, and cell division. The level of c-di-GMP in cells is controlled by diguanyl cyclases (DGCs) and phosphodiesterases (PDEs). Here, we report the biochemical functions and crystal structure of the potential diguanylase Dcsbis (PA2771, a diguanylate cyclase with a self-blocked I-site) from Pseudomonas aeruginosa PAO1. The full-length Dcsbis protein contains an N-terminal GAF domain and a C-terminal GGDEF domain. We showed that Dcsbis tightly coordinates cell motility without markedly affecting biofilm formation and is a diguanylate cyclase with a catalytic activity much higher than those of many other DGCs. Unexpectedly, we found that a peptide loop (protecting loop) extending from the GAF domain occupies the conserved inhibition site, thereby largely relieving the product-inhibition effect. A large hydrophobic pocket was observed in the GAF domain, thus suggesting that an unknown upstream signaling molecule may bind to the GAF domain, moving the protecting loop from the I-site and thereby turning off the enzymatic activity.
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Ying
authors.initials: Y
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Shiheng
authors.initials: S
authors.lastname: Liu
authors.affiliation: Institute for metabolic and neuropsychiatric disorders, Binzhou Medical University, Binzhou, Shandong, 256600, China.
authors.forename: Cuilan
authors.initials: C
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Yan
authors.initials: Y
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Kaikai
authors.initials: K
authors.lastname: Chi
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Tiantian
authors.initials: T
authors.lastname: Su
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Deyu
authors.initials: D
authors.lastname: Zhu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Jin
authors.initials: J
authors.lastname: Peng
authors.affiliation: College of Life Sciences, Shandong Normal University, Jinan, Shandong, 250014, China.
authors.forename: Zhijie
authors.initials: Z
authors.lastname: Xia
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Jing
authors.initials: J
authors.lastname: He
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Sujuan
authors.initials: S
authors.lastname: Xu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.affiliation: State Key Laboratory of Microbial Technology, School of Life Sciences, Shandong University, Jinan, Shandong, 250100, China.
authors.forename: Lichuan
authors.initials: L
authors.lastname: Gu
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000378:metabolism; D018441:Biofilms; D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D006152:Cyclic GMP / Q000031:analogs & derivatives* / Q000378:metabolism; D029968:Escherichia coli Proteins / Q000737:chemistry* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D006860:Hydrogen Bonding; D008958:Models, Molecular; D019761:Phosphorus-Oxygen Lyases / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D000072417:Protein Domains; D011550:Pseudomonas aeruginosa / Q000737:chemistry / Q000201:enzymology / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C062025:bis(3',5')-cyclic diguanylic acid; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase; D006152:Cyclic GMP