The <i>cidA</i> murein hydrolase regulator contributes to DNA release and biofilm development in <i>Staphylococcus aureus</i>

TLDR

cidA and lrgA genes influence cell lysis in *Staphylococcus aureus*, and are thought to encode holin and antiholin proteins that regulate bacterial cell death. The study aimed to determine how cidA‑mediated cell lysis affects biofilm formation in *S. aureus*. Researchers compared biofilm development in a cidA mutant (KB1050) with the parental strain (UAMS‑1) using microscopy, viability staining, qRT‑PCR, and DNase I treatment.

Abstract

The Staphylococcus aureus cidA and lrgA genes have been shown to affect cell lysis under a variety of conditions during planktonic growth. It is hypothesized that these genes encode holins and antiholins, respectively, and may serve as molecular control elements of bacterial cell lysis. To examine the biological role of cell death and lysis, we studied the impact of the cidA mutation on biofilm development. Interestingly, this mutation had a dramatic impact on biofilm morphology and adherence. The cidA mutant (KB1050) biofilm exhibited a rougher appearance compared with the parental strain (UAMS-1) and was less adherent. Propidium iodide staining revealed that KB1050 accumulated more dead cells within the biofilm population relative to UAMS-1, indicative of reduced cell lysis. In agreement with this finding, quantitative real-time PCR experiments demonstrated the presence of 5-fold less genomic DNA in the KB1050 biofilm relative to UAMS-1. Furthermore, treatment of the UAMS-1 biofilm with DNase I caused extensive cell detachment, whereas similar treatment of the KB1050 biofilm had only a modest effect. These results demonstrate that cidA-controlled cell lysis plays a significant role during biofilm development and that released genomic DNA is an important structural component of S. aureus biofilm.