title: Rot is a key regulator of Staphylococcus aureus biofilm formation.
abstract: Staphylococcus aureus is a significant cause of chronic biofilm infections on medical implants. We investigated the biofilm regulatory cascade and discovered that the repressor of toxins (Rot) is part of this pathway. A USA300 community-associated methicillin-resistant S. aureus strain deficient in Rot was unable to form a biofilm using multiple different assays, and we found rot mutants in other strain lineages were also biofilm deficient. By performing a global analysis of transcripts and protein production controlled by Rot, we observed that all the secreted protease genes were up-regulated in a rot mutant, and we hypothesized that this regulation could be responsible for the biofilm phenotype. To investigate this question, we determined that Rot bound to the protease promoters, and we observed that activity levels of these enzymes, in particular the cysteine proteases, were increased in a rot mutant. By inactivating these proteases, biofilm capacity was restored to the mutant, demonstrating they are responsible for the biofilm negative phenotype. Finally, we tested the rot mutant in a mouse catheter model of biofilm infection and observed a significant reduction in biofilm burden. Thus S. aureus uses the transcription factor Rot to repress secreted protease levels in order to build a biofilm.
authors.affiliation: Department of Microbiology, Roy J. and Lucille A. Carver College of Medicine, University of Iowa, Iowa City, IA, 52242, USA.
authors.forename: Joe M
authors.initials: JM
authors.lastname: Mootz
authors.forename: Meredith A
authors.initials: MA
authors.lastname: Benson
authors.forename: Cortney E
authors.initials: CE
authors.lastname: Heim
authors.forename: Heidi A
authors.initials: HA
authors.lastname: Crosby
authors.forename: Jeffrey S
authors.initials: JS
authors.lastname: Kavanaugh
authors.forename: Paul M
authors.initials: PM
authors.lastname: Dunman
authors.forename: Tammy
authors.initials: T
authors.lastname: Kielian
authors.forename: Victor J
authors.initials: VJ
authors.lastname: Torres
authors.forename: Alexander R
authors.initials: AR
authors.lastname: Horswill
mesh_terms: D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D018441:Biofilms; D015964:Gene Expression Regulation, Bacterial; D006801:Humans; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D010447:Peptide Hydrolases / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012097:Repressor Proteins / Q000235:genetics / Q000378:metabolism*; D013203:Staphylococcal Infections / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000235:genetics / Q000502:physiology*
chemical_list: D001426:Bacterial Proteins; D012097:Repressor Proteins; C408643:rot protein, Staphylococcus aureus; D010447:Peptide Hydrolases