Cyclic-di-GMP disodium

• CAS: 2222132-40-1
• Catalog Number: ACM-ONT2222132401
• Category: Cyclic Dinucleotides
• Molecular Weight: 734.37
• Molecular Formula: C20H22N10Na2O14P2

Case Study

Cyclic-di-GMP for Investigating the DgcY/HhmR Regulatory Network in Osmotic Stress Response

Zhang Y, et al. mBio, 2025, 16(9).

Cyclic-di-GMP was applied as a signaling molecule to investigate the regulatory mechanism controlling osmotic stress adaptation in Halomonas hydrothermalis Y2. Researchers constructed an in-frame deletion mutant of the diguanylate cyclase gene dgcY, which encodes a protein containing six transmembrane helices and a GGDEF catalytic domain. Phenotypic assays demonstrated that deletion of dgcY significantly reduced biofilm formation and impaired bacterial growth under high-salt conditions, particularly at low temperatures. To elucidate the regulatory mechanism, protein interaction assays were performed between DgcY and the LysR-type transcriptional regulator HhmR. Biochemical analysis revealed that cyclic-di-GMP promotes the formation of a DgcY-HhmR complex, resulting in feedback inhibition of DgcY catalytic activity. Additional hhmR knockout experiments combined with motility and biofilm assays further confirmed the functional role of this regulatory interaction. These results demonstrate that cyclic-di-GMP serves as a key experimental probe for dissecting feedback regulatory mechanisms governing bacterial adaptation to osmotic stress.

Cyclic-di-GMP for Regulation Analysis of NadR-Mediated NAD Biosynthesis in Salmonella

Mao L, et al. mBio, 2025, 16(9), 2025.

Cyclic di-GMP was employed to examine its regulatory role in NAD metabolism in Salmonella enterica serovar Typhimurium. A transcription factor binding screen was conducted to identify cyclic-di-GMP-responsive proteins, leading to the discovery of NadR, a transcriptional repressor controlling genes involved in NAD synthesis and salvage pathways. In vitro binding assays demonstrated that cyclic-di-GMP interacts with NadR with high affinity. Electrophoretic DNA-binding analyses showed that cyclic-di-GMP binding inhibits the association of NadR with its target promoter sequences, thereby derepressing NAD biosynthetic genes. Enzymatic activity assays further revealed that cyclic-di-GMP enhances NadR-associated nicotinamide mononucleotide adenylyltransferase and ribosylnicotinamide kinase activities. Cellular experiments confirmed that elevated intracellular cyclic-di-GMP levels increase NAD production and improve bacterial resistance to DNA damage. This study highlights the use of cyclic-di-GMP as a biochemical regulator to elucidate transcriptional and metabolic control mechanisms governing bacterial redox homeostasis.

Cyclic-di-GMP for Functional Analysis of Virulence Regulation in Pseudomonas syringae

Sun X, et al. Journal of Invertebrate Pathology, 2024, 207, 108189.

Cyclic di-GMP was experimentally manipulated to investigate its role in virulence regulation of Pseudomonas syringae during Caenorhabditis elegans infection. Recombinant plasmids were engineered to modulate intracellular cyclic-di-GMP levels: plasmid pDGC, carrying a GGDEF-domain diguanylate cyclase gene (VT47_20995), was introduced to elevate cyclic-di-GMP synthesis, whereas plasmid pPDE, encoding an EAL-domain phosphodiesterase gene, was used to reduce cyclic-di-GMP levels. These constructs were transformed into the wild-type MB03 strain and the fleQ03 knockout mutant to evaluate the regulatory role of the FleQ transcription factor. GFP-labeled bacterial strains were utilized to visualize infection dynamics in C. elegans. Comparative infection assays under controlled antibiotic selection conditions enabled precise assessment of virulence phenotypes associated with altered cyclic-di-GMP signaling. This experimental design demonstrates how cyclic-di-GMP-modulating genetic systems can be used to dissect bacterial signaling pathways controlling host-pathogen interactions.