Cyclic di-GMP Signaling in Bacillus subtilis Is Governed by Direct Interactions of Diguanylate Cyclases and Cognate Receptors.

MBio
Sandra KunzPeter L Graumann

Abstract

Bacillus subtilis contains two known cyclic di-GMP (c-di-GMP)-dependent receptors, YdaK and DgrA, as well as three diguanylate cyclases (DGCs): soluble DgcP and membrane-integral DgcK and DgcW. DgrA regulates motility, while YdaK is responsible for the formation of a putative exopolysaccharide, dependent on the activity of DgcK. Using single-molecule tracking, we show that a majority of DgcK molecules are statically positioned in the cell membrane but significantly less so in the absence of YdaK but more so upon overproduction of YdaK. The soluble domains of DgcK and of YdaK show a direct interaction in vitro, which depends on an intact I-site within the degenerated GGDEF domain of YdaK. These experiments suggest a direct handover of a second messenger at a single subcellular site. Interestingly, all three DGC proteins contribute toward downregulation of motility via the PilZ protein DgrA. Deletion of dgrA also affects the mobility of DgcK within the membrane and also that of DgcP, which arrests less often at the membrane in the absence of DgrA. Both, DgcK and DgcP interact with DgrA in vitro, showing that divergent as well as convergent direct connections exist between cyclases and their effector proteins. Automated determinat...Continue Reading

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Citations

Nov 15, 2020·NPJ Biofilms and Microbiomes·Anne BoyeldieuCécile Jourlin-Castelli
Mar 1, 2021·Trends in Microbiology·Regine Hengge
May 18, 2021·Frontiers in Microbiology·Benjamin MayerPeter L Graumann
Jun 8, 2021·Journal of Bacteriology·Timothy M HermanasGeorge C Stewart

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Datasets Mentioned

BETA
GMQE

Methods Mentioned

BETA
pulldown
pull-down
fluorescence microscopy
motility assays
PCR
size exclusion chromatography

Software Mentioned

SWISS
MATLAB
MODEL
VisiView
ImageJ
Fiji
ProteinLynx Global Server
SMTracker
SMM Track
MEMHDX

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