Impact of the regulatory loci agr, sarA and sae of Staphylococcus aureus on the induction of alpha-toxin during device-related infection resolved by direct quantitative transcript analysis.
The cytotoxic alpha-toxin (encoded by hla) of Staphylococcus aureus is regulated by three loci, agr, sarA and sae, in vitro. Here, we assess the regulation of hla in a guinea pig model of device-related infection by quantifying RNAIII (the effector molecule of agr) and hla directly in exudates accumulating in infected devices without subculturing of the bacteria. LightCycler reverse transcription-polymerase chain reaction (RT-PCR) was used to quantify the transcripts. Strains RN6390 and Newman expressed considerably smaller amounts of RNAIII in the guinea pig than during in vitro growth. The residual RNAIII expression decreased during the course of infection and was negatively correlated with bacterial densities. As with RNAIII, the highest hla expression was detected in both strains early in infection. Even in strain Newman, a weak hla producer in vitro, a pronounced expression of hla was observed during infection. Likewise, four S. aureus isolates from cystic fibrosis (CF) patients expressed Q1hla despite an inactive agr during device-related infection as in the CF lung. Mutation of agr and sarA in strain Newman and RN6390 had no consequence for hla expression in vivo. In contrast, the mutation in sae resulted in severe downregulation of hla in vitro as well as in vivo. In conclusion, S. aureus seems to be provided with regulatory circuits different from those characterized in vitro to ensure alpha-toxin synthesis during infections.
Goerke
C
C
Institute for General and Environmental Hygiene, University of Tübingen, Wilhelmstrasse 31, 72074 Tübingen, Germany. christiane.wolz@uni-tuebingen.de
Fluckiger
U
U
Steinhuber
A
A
Zimmerli
W
W
Wolz
C
C
D000818:Animals; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D001427:Bacterial Toxins / Q000235:genetics / Q000378:metabolism*; D003550:Cystic Fibrosis / Q000382:microbiology; D005122:Exudates and Transudates; D015964:Gene Expression Regulation, Bacterial; D005809:Genes, Regulator; D006168:Guinea Pigs; D006460:Hemolysin Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D009154:Mutation; D016459:Prosthesis-Related Infections / Q000235:genetics / Q000382:microbiology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013203:Staphylococcal Infections / Q000235:genetics / Q000382:microbiology*; D013211:Staphylococcus aureus / Q000235:genetics* / Q000472:pathogenicity*; D015534:Trans-Activators; D014157:Transcription Factors
D001426:Bacterial Proteins; D001427:Bacterial Toxins; D006460:Hemolysin Proteins; C433805:SaeR protein, Staphylococcus aureus; C088021:SarA protein, bacterial; D015534:Trans-Activators; D014157:Transcription Factors; C021091:staphylococcal alpha-toxin