Novel TPP-riboswitch activators bypass metabolic enzyme dependency

Area of Science:

• Molecular Biology
• Biochemistry
• Microbiology

Background:

• Riboswitches are mRNA regulatory elements controlling gene expression.
• TPP-riboswitches regulate thiamine metabolism and are potential antibacterial targets.
• Thiamine metabolism is essential for bacterial survival.

Purpose of the Study:

• To develop novel thiamine analogs targeting TPP-riboswitches.
• To investigate the mechanism of action and antibacterial potential of these analogs.
• To explore compound activation dependence on metabolic proteins.

Main Methods:

• Synthesis of thiamine analogs with a 1,2,3-triazole group.
• Reporter gene assays in E. coli strains to measure riboswitch repression.
• Investigating the role of thiamine kinase (ThiK) in compound activation.
• Testing triazolethiamine derivatives with phosphate-mimicking moieties.

Main Results:

• Thiamine analogs, particularly triazolethiamine (TT), repressed thiM-riboswitch gene expression.
• Compound activation was dependent on proteins involved in thiamine metabolism, like ThiK.
• Triazolethiamine showed higher activity than pyrithiamine (PT).
• Phosphate-mimicking derivatives bypassed ThiK-dependent activation.

Conclusions:

• Triazolethiamine is a potent TPP-riboswitch modulator with superior activity to existing compounds.
• Metabolic activation is crucial for some riboswitch-targeting compounds.
• Novel antibacterial agents targeting TPP-riboswitches can be developed.
• Design of compounds independent of endogenous activation mechanisms is feasible.