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Bacteria possess a repertoire of distinct regulatory systems promoting survival in disparate environments. Under in vitro conditions it was demonstrated for the human pathogenStaphylococcus aureus that the expression of most virulence factors is coordinated by the global regulatoragr . To monitor bacterial gene regulation in the host, we developed a method for direct transcript analysis from clinical specimens. Quantification of specific transcripts was performed by competitive reverse transcription-PCR, and results were normalized against the constitutively expressed gene for gyrase (gyr ). Using sputum from cystic fibrosis (CF) patients infected withS. aureus we examined the transcription of the effector molecule RNAIII ofagr , ofspa (protein A), generally repressed byagr , and ofhla (alpha-toxin), generally activated byagr . In the CF lung RNAIII was expressed poorly, indicating an inactiveagr in vivo. Despite the low level of RNAIII expression,spa was detectable only in minute amounts and an irregular transcription ofhla was observed in all sputum samples. After subculturing of patient strainsagr -deficient isolates and isolates with unusual expression profiles, i.e., not consistent with those obtained from prototypic strains, were observed. In conclusion, theagr activity seems to be nonessential in CF, and from the described expression pattern ofspa andhla , other regulatory circuits aside fromagr are postulated in vivo.

infection and immunity

Direct Quantitative Transcript Analysis of the <i>agr</i> Regulon of <i>Staphylococcus aureus</i> during Human Infection in Comparison to the Expression Profile In Vitro

Direct Quantitative Transcript Analysis of the agr Regulon of Staphylococcus aureus during Human Infection in Comparison to the Expression Profile In Vitro