Proteome Dynamics during Antibiotic Persistence and Resuscitation

Area of Science:

• Microbiology
• Molecular Biology
• Biochemistry

Background:

• Antibiotic persister cells are transiently tolerant to antibiotics by reducing growth and metabolism.
• Characterizing persister cells is challenging due to their low numbers and need for isolation.
• Toxin-antitoxin systems, like hipA/hipB in Escherichia coli, are key in inducing persistence and resuscitation.

Purpose of the Study:

• To develop a method for temporal analysis of protein synthesis during antibiotic persistence and resuscitation.
• To identify newly synthesized proteins in viable persister cells.
• To understand the molecular mechanisms underlying bacterial survival and recovery from antibiotic treatment.

Main Methods:

• Utilized stable isotope-labeled lysine for proteome labeling during hipA-induced persistence and hipB-induced resuscitation in Escherichia coli.
• Performed time-resolved, 24-hour measurements of label incorporation.
• Analyzed over 500 newly synthesized proteins in viable cells.

Main Results:

• Demonstrated low but widespread protein synthesis during antibiotic persistence.
• Identified essential proteins and ribosome-associated proteins (RaiA, Sra) synthesized during persistence, suggesting roles in survival.
• Observed synthesis of ribosome-splitting GTPase HflX and ABC transporters during resuscitation, alongside restored translation and metabolism (glycolysis, amino acid biosynthesis).

Conclusions:

• Provides the first global analysis of protein synthesis in bacterial persister and resuscitating cells.
• Highlights the dynamic proteome changes governing antibiotic persistence and recovery.
• Offers a valuable resource for studying the molecular processes of antibiotic tolerance and bacterial survival.