title: Staphylococcus aureus CidA and LrgA proteins exhibit holin-like properties.
abstract: The Staphylococcus aureus cid and lrg operons are known to be involved in biofilm formation by controlling cell lysis and the release of genomic DNA, which ultimately becomes a structural component of the biofilm matrix. Although the molecular mechanisms controlling cell death and lysis are unknown, it has been hypothesized that the cidA and lrgA genes encode holin- and antiholin-like proteins and function to regulate these processes similarly to bacteriophage-induced death and lysis. In this study, we focused on the biochemical and molecular characterization of CidA and LrgA with the goal of testing the holin model. First, membrane fractionation and fluorescent protein fusion studies revealed that CidA and LrgA are membrane-associated proteins. Furthermore, similarly to holins, CidA and LrgA were found to oligomerize into high-molecular-mass complexes whose formation was dependent on disulfide bonds formed between cysteine residues. To determine the function of disulfide bond-dependent oligomerization of CidA, an S. aureus mutant in which the wild-type copy of the cidA gene was replaced with the cysteine mutant allele was generated. As determined by β-galactosidase release assays, this mutant exhibited increased cell lysis during stationary phase, suggesting that oligomerization has a negative impact on this process. When analyzed for biofilm development and maturation, this mutant displayed increased biofilm adhesion in a static assay and a greater amount of dead-cell accumulation during biofilm maturation. These studies support the model that CidA and LrgA proteins are bacterial holin-/antiholin-like proteins that function to control cell death and lysis during biofilm development.
authors.affiliation: Department of Microbiology and Immunology, University of Arkansas for Medical Sciences, Little Rock, Arkansas 72205, USA.
authors.forename: Dev K
authors.initials: DK
authors.lastname: Ranjit
authors.forename: Jennifer L
authors.initials: JL
authors.lastname: Endres
authors.forename: Kenneth W
authors.initials: KW
authors.lastname: Bayles
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D001433:Bacteriolysis; D018441:Biofilms / Q000254:growth & development; D002462:Cell Membrane / Q000737:chemistry; D002463:Cell Membrane Permeability / Q000502:physiology; D008565:Membrane Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D050296:Microbial Viability; D055503:Protein Multimerization; D013211:Staphylococcus aureus / Q000737:chemistry / Q000201:enzymology* / Q000254:growth & development / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; C406202:LrgA protein, Staphylococcus aureus; D008565:Membrane Proteins