title: CcpA-dependent carbohydrate catabolite repression regulates galactose metabolism in Streptococcus oligofermentans.
abstract: Streptococcus oligofermentans is an oral commensal that inhibits the growth of the caries pathogen Streptococcus mutans by producing copious amounts of H(2)O(2) and that grows faster than S. mutans on galactose. In this study, we identified a novel eight-gene galactose (gal) operon in S. oligofermentans that was comprised of lacABCD, lacX, and three genes encoding a galactose-specific transporter. Disruption of lacA caused more growth reduction on galactose than mutation of galK, a gene in the Leloir pathway, indicating that the principal role of this operon is in galactose metabolism. Diauxic growth was observed in cultures containing glucose and galactose, and a luciferase reporter fusion to the putative gal promoter demonstrated 12-fold repression of the operon expression by glucose but was induced by galactose, suggesting a carbon catabolite repression (CCR) control in galactose utilization. Interestingly, none of the single-gene mutations in the well-known CCR regulators ccpA and manL affected diauxic growth, although the operon expression was upregulated in these mutants in glucose. A double mutation of ccpA and manL eliminated glucose repression of galactose utilization, suggesting that these genes have parallel functions in regulating gal operon expression and mediating CCR. Electrophoretic mobility shift assays demonstrated binding of CcpA to the putative catabolite response element motif in the promoter regions of the gal operon and manL, suggesting that CcpA regulates CCR through direct regulation of the transcription of the gal operon and manL. This provides the first example of oral streptococci using two parallel CcpA-dependent CCR pathways in controlling carbohydrate metabolism.
authors.affiliation: State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
authors.forename: Jun
authors.initials: J
authors.lastname: Cai
authors.forename: Huichun
authors.initials: H
authors.lastname: Tong
authors.forename: Fengxia
authors.initials: F
authors.lastname: Qi
authors.forename: Xiuzhu
authors.initials: X
authors.lastname: Dong
mesh_terms: D001665:Binding Sites; D057465:Catabolite Repression; D003470:Culture Media / Q000737:chemistry; D004269:DNA, Bacterial / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D005690:Galactose / Q000378:metabolism*; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D015971:Gene Expression Regulation, Enzymologic; D005798:Genes, Bacterial; D017930:Genes, Reporter; D005947:Glucose / Q000378:metabolism; D008156:Luciferases / Q000032:analysis / Q000235:genetics; D009876:Operon; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D013291:Streptococcus / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism*
chemical_list: D003470:Culture Media; D004269:DNA, Bacterial; D004268:DNA-Binding Proteins; D008156:Luciferases; D005947:Glucose; D005690:Galactose