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PKA was induced in HeLa 229 cells by treatment with 0.1 mM isobutylmethylxanthine for 16 h or with 0.2 mM forskolin for 24 h. To determine if PKA is a relevant kinase for BLA regulation, we infected HeLa 229 cells with C. trachomatis and then treated them with either 0.1 mM isobutylmethylxanthine or 0.2 mM forskolin for 16 h. The amounts of BLA and PKA proteins were determined by immunoblotting and radioimmunoprecipitation assays. The cellular localization of BLA was determined by confocal laser scanning microscopy and by in situ hybridization. Beta-lactamase activity was measured by colorimetric assay. The results showed that treatment of C. trachomatis-infected cells with PKA activators reduced BLA secretion to undetectable levels. In contrast, inhibitors of PKA prevented the decrease in BLA secretion caused by C. trachomatis infection. Moreover, the PKA inhibitor H89 reduced BLA activity but had no effect on BLA protein expression. 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