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Characterization of RAT, an autolysis regulator in Staphylococcus aureus.

S S Ingavale¹, W Van Wamel, A L Cheung

¹Department of Microbiology, Dartmouth Medical School, Hanover, NH 03755, USA.

Molecular Microbiology

|June 7, 2003

Summary

Researchers identified a new gene, regulator of autolytic activity (rat), in Staphylococcus aureus. This gene controls bacterial cell wall breakdown, impacting growth and antibiotic sensitivity.

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Area of Science:

Microbiology
Molecular Biology
Genetics

Background:

Staphylococcus aureus pathogenesis involves complex regulatory networks controlling virulence factors.
Autolytic activity, crucial for cell wall remodeling and bacterial life cycle, is tightly regulated.
Understanding these regulatory mechanisms is key to developing novel therapeutic strategies.

Purpose of the Study:

To identify and characterize novel genetic regulators of autolytic activity in Staphylococcus aureus.
To elucidate the function of a newly discovered gene, designated rat (regulator of autolytic activity).

Main Methods:

Transposon mutagenesis to identify mutants with altered phenotypes.
Sequence analysis to identify the disrupted gene.
Gene expression analysis (e.g., Northern blotting, qRT-PCR) to assess target gene regulation.

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Gel shift assays to confirm DNA-binding activity of the Rat protein.

Main Results:

A transposon mutant displayed growth defects, enhanced autolysis, and increased sensitivity to Triton X-100 and penicillin.
The disrupted gene, named rat, encodes a 147-amino acid protein homologous to MarR and SarA families.
Mutations in rat led to decreased expression of autolytic regulators lytSR, lrgAB, and arlRS.
Rat protein directly binds to the promoters of lytRS and arlRS, acting as a repressor.
Rat negatively regulates autolysin genes, including lytM and lytN.

Conclusions:

The rat gene product is a significant negative regulator of autolytic activity in Staphylococcus aureus.
Rat plays a crucial role in controlling bacterial autolysis, influencing growth and antibiotic susceptibility.
Rat represents a potential target for therapeutic intervention in Staphylococcus aureus infections.