Phosphorylation-independent regulation of the diguanylate cyclase WspR.

PLoS biology (2008-03-28)
Nabanita De, Michelle Pirruccello, Petya Violinova Krasteva, Narae Bae, Rahul Veera Raghavan, Holger Sondermann
ABSTRACT

Environmental signals that trigger bacterial pathogenesis and biofilm formation are mediated by changes in the level of cyclic dimeric guanosine monophosphate (c-di-GMP), a unique eubacterial second messenger. Tight regulation of cellular c-di-GMP concentration is governed by diguanylate cyclases and phosphodiesterases, which are responsible for its production and degradation, respectively. Here, we present the crystal structure of the diguanylate cyclase WspR, a conserved GGDEF domain-containing response regulator in Gram-negative bacteria, bound to c-di-GMP at an inhibitory site. Biochemical analyses revealed that feedback regulation involves the formation of at least three distinct oligomeric states. By switching from an active to a product-inhibited dimer via a tetrameric assembly, WspR utilizes a novel mechanism for modulation of its activity through oligomerization. Moreover, our data suggest that these enzymes can be activated by phosphodiesterases. Thus, in addition to the canonical pathways via phosphorylation of the regulatory domains, both product and enzyme concentration contribute to the coordination of c-di-GMP signaling. A structural comparison reveals resemblance of the oligomeric states to assemblies of GAF domains, widely used regulatory domains in signaling molecules conserved from archaea to mammals, suggesting a similar mechanism of regulation.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, fatty acid free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, essentially globulin free, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, fatty acid free, low endotoxin, lyophilized powder, BioReagent, suitable for cell culture, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, cold ethanol fraction, pH 5.2, ≥96%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially fatty acid free, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, suitable for RIA, pH 5.2, ≥96%
Sigma-Aldrich
Congo Red, Dye content ≥35 %
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 5.2, ≥96%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, lyophilized powder, essentially fatty acid free, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, suitable for (for molecular biology), Non-acetylated
Sigma-Aldrich
Bovine Serum Albumin, pH <5.0, powder
Sigma-Aldrich
Bovine Serum Albumin, powder, BioXtra
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, suitable for hybridization, pH 7, ≥98%
Sigma-Aldrich
Congo Red, certified by the Biological Stain Commission, BioXtra
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially globulin free, ≥99% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially IgG-free, low endotoxin, BioReagent, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially fatty acid free and essentially globulin free, ≥99% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, low endotoxin, lyophilized powder, Cohn Analog, BioReagent, suitable for cell culture, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, Cohn Fraction V, lyophilized powder, ≥96% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, pH 5.2, ≥96%
Sigma-Aldrich
Bovine Serum Albumin, heat shock fraction, protease free, low endotoxin, suitable for cell culture, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, chromatographically purified, New Zealand origin, low endotoxin, suitable for cell culture, pH 7, ≥98%
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially globulin free, BioReagent, suitable for cell culture
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, essentially globulin free, low endotoxin, ≥98% (agarose gel electrophoresis)
Sigma-Aldrich
Bovine Serum Albumin, lyophilized powder, ≥98% (agarose gel electrophoresis)