CcpA-dependent and -independent control of beta-galactosidase expression in Streptococcus pneumoniae occurs via regulation of an upstream phosphotransferase system-encoding operon.
A spontaneous mutant of Streptococcus pneumoniae strain D39 exhibiting elevated beta-galactosidase activity was identified. We determined that the beta-galactosidase activity was due to BgaA, a surface protein in S. pneumoniae, and that the expression of bgaA was regulated. Transcription analyses demonstrated expression of bgaA in the constitutive beta-galactosidase (BgaA(C)) mutant, but not in the parent. beta-Galactosidase expression was induced in the parent under specific growth conditions; however, the levels did not reach those of the BgaA(C) mutant. We localized the mutation resulting in the BgaA(C) phenotype to a region upstream of bgaA and in the promoter of a phosphoenolpyruvate-dependent phosphotransferase system (PTS) operon. The mutation was in a catabolite-responsive element (cre) and affected the binding of CcpA (catabolite control protein A), a key regulator of many carbon metabolism genes. The pts operon and bgaA were cotranscribed, and their transcription was regulated by CcpA. Deletion of ccpA altered beta-galactosidase activity, leading to a sevenfold increase in the parent but a fivefold decrease in the BgaA(C) mutant. The resulting beta-galactosidase activities were the same in the two strains, suggesting the presence of a second repressor. The presence of glucose in the growth medium resulted in pts-bgaA repression by both CcpA and the second repressor, with the latter being important in responding to the glucose concentration. Expression of beta-galactosidase is important for S. pneumoniae adherence during colonization of the nasopharynx, a site normally devoid of glucose. CcpA and environmental glucose concentrations thus appear to play important roles in the regulation of a niche-specific virulence factor.
Kaufman
Greer E
GE
Department of Microbiology, Univerisity of Alabama at Birmingham, Birmingham, AL 35294-2170, USA.
Yother
Janet
J
D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D024202:Electrophoretic Mobility Shift Assay; D017353:Gene Deletion; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D008969:Molecular Sequence Data; D009154:Mutation; D009876:Operon; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D012097:Repressor Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D013296:Streptococcus pneumoniae / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D014158:Transcription, Genetic; D001616:beta-Galactosidase / Q000235:genetics* / Q000378:metabolism
D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D012097:Repressor Proteins; C069286:catabolite control proteins, bacteria; D010731:Phosphoenolpyruvate Sugar Phosphotransferase System; D001616:beta-Galactosidase; D005947:Glucose