title: Structural basis of activity and allosteric control of diguanylate cyclase.
abstract: Recent discoveries suggest that a novel second messenger, bis-(3'-->5')-cyclic di-GMP (c-diGMP), is extensively used by bacteria to control multicellular behavior. Condensation of two GTP to the dinucleotide is catalyzed by the widely distributed diguanylate cyclase (DGC or GGDEF) domain that occurs in various combinations with sensory and/or regulatory modules. The crystal structure of the unorthodox response regulator PleD from Caulobacter crescentus, which consists of two CheY-like receiver domains and a DGC domain, has been solved in complex with the product c-diGMP. PleD forms a dimer with the CheY-like domains (the stem) mediating weak monomer-monomer interactions. The fold of the DGC domain is similar to adenylate cyclase, but the nucleotide-binding mode is substantially different. The guanine base is H-bonded to Asn-335 and Asp-344, whereas the ribosyl and alpha-phosphate moieties extend over the beta2-beta3-hairpin that carries the GGEEF signature motif. In the crystal, c-diGMP molecules are crosslinking active sites of adjacent dimers. It is inferred that, in solution, the two DGC domains of a dimer align in a two-fold symmetric way to catalyze c-diGMP synthesis. Two mutually intercalated c-diGMP molecules are found tightly bound at the stem-DGC interface. This allosteric site explains the observed noncompetitive product inhibition. We propose that product inhibition is due to domain immobilization and sets an upper limit for the concentration of this second messenger in the cell.
authors.affiliation: Divisions of Structural Biology and Molecular Microbiology, Biozentrum, University of Basel, Klingelbergstrasse 70, 4056 Basel, Switzerland.
authors.forename: Carmen
authors.initials: C
authors.lastname: Chan
authors.forename: Ralf
authors.initials: R
authors.lastname: Paul
authors.forename: Dietrich
authors.initials: D
authors.lastname: Samoray
authors.forename: Nicolas C
authors.initials: NC
authors.lastname: Amiot
authors.forename: Bernd
authors.initials: B
authors.lastname: Giese
authors.forename: Urs
authors.initials: U
authors.lastname: Jenal
authors.forename: Tilman
authors.initials: T
authors.lastname: Schirmer
mesh_terms: D000494:Allosteric Regulation; D001426:Bacterial Proteins / Q000737:chemistry* / Q000378:metabolism; D001665:Binding Sites; D002384:Catalysis; D016935:Caulobacter crescentus / Q000201:enzymology*; D018360:Crystallography, X-Ray; D029968:Escherichia coli Proteins; D008958:Models, Molecular; D015394:Molecular Structure; D019761:Phosphorus-Oxygen Lyases / Q000737:chemistry* / Q000378:metabolism; D011485:Protein Binding; D011487:Protein Conformation
chemical_list: D001426:Bacterial Proteins; D029968:Escherichia coli Proteins; C096455:PleD protein, Caulobacter crescentus; D019761:Phosphorus-Oxygen Lyases; C060720:diguanylate cyclase