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Detection and characterization of cyclic di-GMP-binding proteins

Project description

Identifying and characterising c-di-GMP binding receptors in bacteria

Cyclic di-GMP (c-di-GMP) is a bacterial ‘second messenger’ molecule that plays a crucial role in regulating a wide range of cellular processes, including biofilm formation, motility, cell cycle progression and virulence. Receptors that bind c-di-GMP are essential for mediating these intracellular responses. However, their diversity makes them challenging to characterise and thus impedes deeper understanding complex regulation of microbial pathways that contribute to the exceptional adaptation and evolution of bacteria. With the support of the Marie Skłodowska-Curie Actions programme, the CdiGMPBP project aims to leverage a novel screen for c-di-GMP binding proteins, based on alterations in thermal stability, to fill important gaps in knowledge.

Objective

Nucleotide signaling plays an important role in the adjustment of microbial physiology and metabolism. These signaling systems are involved in the regulation of all physiological processes including biofilm formation, virulence, environmental survival, and phage defense. One of the most prominent nucleotide-based second messenger molecules is the ubiquitous second messenger cyclic di-GMP. Cyclic di‐GMP synthesizing and hydrolyzing proteins, GGDEF and EAL domain proteins, respectively, are widely present across bacterial phyla, numerous within genomes, and highly conserved throughout the phylogenetic tree. However, there is still limited understanding of c-di-GMP binding receptors, since they are diverse, not necessarily characterized by common signature binding motifs, and still require experimentation to be identified.

Having established a novel screen for cyclic di-GMP binding proteins based on alterations in thermal stability (PISA), this project aims to identify and characterize novel c-di-GMP binding receptors of the gastrointestinal pathogen Salmonella typhimurium as a model organism. Candidate c-di-GMP binding proteins will be verified by alternative approaches. Verified receptors will be assessed using microbiology, biochemistry, molecular biology, bioinformatics, and genetic engineering approaches. While new cyclic di-GMP binding proteins will be predominantly characterized in Salmonella typhimurium, novel and improved screens for cyclic di-GMP binding proteins based on mass-spectrometry-based DRaCALA and isolation of cellular nanomachines like ribosomes will be developed and partially investigated as research resources. Successful conduction of this post-doctoral project will expand the number of c-di-GMP receptors, discover novel underlying mechanisms of regulation by c-di-GMP, and thus significantly improve our understanding of the multilayer regulation of microbial pathways that contribute to the exceptional adaptation and evolution of bacteria.

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HORIZON-TMA-MSCA-PF-EF - HORIZON TMA MSCA Postdoctoral Fellowships - European Fellowships

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Call for proposal

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(opens in new window) HORIZON-MSCA-2024-PF-01

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Coordinator

KAROLINSKA INSTITUTET
Net EU contribution

Net EU financial contribution. The sum of money that the participant receives, deducted by the EU contribution to its linked third party. It considers the distribution of the EU financial contribution between direct beneficiaries of the project and other types of participants, like third-party participants.

€ 236 340,00
Address
NOBELS VAG 5
171 77 STOCKHOLM
Sweden

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Region
Östra Sverige Stockholm Stockholms län
Activity type
Higher or Secondary Education Establishments
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Total cost

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