Unusual features and molecular pathways of Staphylococcus aureus L-form bacteria

Staphylococcus aureus can be converted to cell wall-deficient L-form bacteria in specific environment which is associated with recurrent and persistent infections. The biophysical properties and molecular basis involved in S. aureus L-form formation are poorly understood. Here, S. aureus unstable L-form model was established not only in Newman strain, but also in ATCC 25923 and five different antibiotic-resistant clinical strains, and the morphology and mechanical properties of Newman strain L-forms were characterized by using atomic force microscopy. Meanwhile, zeta potential, growth and proliferation properties, and hemolysis of L-forms were determined. Gene expression changes involved in transition from S. aureus wild type into L-forms were identified. Our studies showed that L-form S. aureus presented pleomorphism, rough surface, and higher elasticity modulus. L-forms were characterized by less surface charge and had higher hemolysis than the walled form. The S. aureus L-form "fried egg" colony was derived from a single bacterium rather than from aggregation of different bacterial cells. Transcriptomics analysis revealed that several pathways involved in energy metabolism, stress response, protein synthesis, RNA metabolism, and virulence were involved in L-form formation in S. aureus. Our results shed new light on the biological properties and mechanisms underlying L-form formation in S. aureus. These findings will not only be useful for understanding the unique properties and mechanisms of L-form bacteria, but also provide therapeutic targets for developing more effective treatments for S. aureus L-forms.