In low-G+C Gram-positive bacteria, the regulatory protein CcpA has been shown to play a major part in the so-called carbon catabolite repression (CCR) process, as well as in the induction of basic metabolic genes, for which it is considered a global regulator. A strain of Lactobacillus casei that carried a complete deletion of ccpA has been constructed and used to test the effect of CCR on N-acetylglucosaminidase activity and growth performance of a collection of seven CcpA mutations obtained by site-directed mutagenesis. The replaced amino acids were located in the DNA- and cofactor (P-Ser-HPr)-binding domains. Mutations in the DNA-binding domain lacked CCR, as found in Bacillus megaterium. However, mutations in the cofactor-binding domain of L. casei CcpA had a different phenotype to that observed in the previous studies with B. megaterium. Two of them, S80L and T307I, displayed a significant hyper-repression, an effect never reported before for CcpA. Comparison of growth capabilities provided by the different mutants and their ability to sustain CCR demonstrated that CCR, at least on the enzymic activity tested, and the growth defect caused by the CcpA mutations are unrelated features.
Aung-HilbrichL. M., SeidelG., WagnerA., HillenW.2002; Quantification of the influence of HPrSer46P on CcpA-cre interaction. J Mol Biol 319:77–85[CrossRef]
BrücknerR., TitgemeyerF.2002; Carbon catabolite repression in bacteria: choice of the carbon source and autoregulatory limitation of sugar utilization. FEMS Microbiol Lett 209:141–148[CrossRef]
DossonnetV., MonederoV., ZagorecM., GalinierA., Pérez-MartínezG., DeutscherJ.2000; Phosphorylation of HPr by the bifunctional HPr Kinase/P-ser-H phosphatase from Lactobacillus casei controls catabolite repression and inducer exclusion but not inducer expulsion. J Bacteriol 182:2582–2590[CrossRef]
EgeterO., BrücknerR.1996; Catabolite repression mediated by the catabolite control protein CcpA in Staphylococcus xylosus. Mol Microbiol 21:739–749[CrossRef]
FriedmanA. M., FischmannT. O., SteitzT. A.1995; Crystal structure of lac repressor core tetramer and its implications for DNA looping. Science 268:1721–1727[CrossRef]
FujitaY., MiwaY., GalinierA., DeutscherJ.1995; Specific recognition of the Bacillus subtilis gntcis-acting catabolite-responsive element by a protein complex formed between CcpA and seryl-phosphorylated HPr. Mol Microbiol 17:953–960[CrossRef]
GiammarinaroP., PatonJ. C.2002; Role of RegM, a homologue of the catabolite repressor protein CcpA, in the virulence of Streptococcus pneumoniae. Infect Immun 70:5454–5461[CrossRef]
GordonA. J., BurnsP. A., FixD. F.7 other authors1988; Missense mutation in the lacI gene of Escherichia coli. Inferences on the structure of the repressor protein. J Mol Biol 200:239–251[CrossRef]
GosseringerR., KüsterE., GalinierA., DeutscherJ., HillenW.1997; Cooperative and non-cooperative DNA binding modes of catabolite control protein CcpA from Bacillus megaterium result from sensing two different signals. J Mol Biol 266:665–676[CrossRef]
HueckC. J., KrausA., SchmiedelD., HillenW.1995; Cloning, expression and functional analyses of the catabolite control protein CcpA from Bacillus megaterium. Mol Microbiol 16:855–864[CrossRef]
JonesB. E., DeutscherJ., KlevitR. E., DossonnetV., KüsterE., HillenW.1997; Binding of the catabolite repressor protein CcpA to its DNA target is regulated by phosphorylation of its corepressor HPr. J Biol Chem 272:26530–26535[CrossRef]
KimJ. H., VoskuilM. I., ChamblissG. H.1998; NADP, corepressor for the Bacillus catabolite control protein CcpA. Proc Natl Acad Sci U S A 95:9590–9595[CrossRef]
KimH. J., RouxA., SonensheinA. L.2002; Direct and indirect roles of CcpA in regulation of Bacillus subtilis Krebs cycle genes. Mol Microbiol 45:179–190[CrossRef]
KleinaL. G., MillerJ. H.1990; Genetic studies of the lac repressor. XIII. Extensive amino acid replacements generated by the use of natural and synthetic nonsense suppressors. J Mol Biol 212:295–318[CrossRef]
KrausA., KüsterE., WagnerA., HoffmannK., HillenW.1998; Identification of a co-repressor binding site in catabolite control protein CcpA. Mol Microbiol 30:955–963[CrossRef]
KüsterE., LuesinkE. J., de VosW. M., HillenW.1996; Immunological crossreactivity to catabolite control protein CcpA from Bacillus megaterium is found in many Gram-positive bacteria. FEMS Microbiol Lett 139:109–115[CrossRef]
KüsterE., HilbichT., DahlM. K., HillenW.1999a; Mutations in catabolite control protein CcpA separating growth effects from catabolite repression. J Bacteriol 181:4125–4128
KüsterE., WagnerA., VölkerU., HillenW.1999b; Mutations in catabolite control protein CcpA showing glucose-independent regulation in Bacillus megaterium. J Bacteriol 181:7634–7638
LewisM., ChangG., HortonN. C., KercherM. A., PaceH. C., SchumacherM. A., BrennanR. G., LuP.1996; Crystal structure of the lactose operon repressor and its complexes with DNA and inducer. Science 271:1247–1254[CrossRef]
LokmanB. C., HeerikhuisenM., BorsboomY., ChaillouS., PostmaP. W., PouwelsP. H., LeerR. J., van den BroekA.1997; Regulation of expression of the Lactobacillus pentosus xylAB operon. J Bacteriol 179:5391–5397
LudwigH., HomuthG., SchmalischM., DykaF. M., HeckerM., StülkeJ.2001; Transcription of glycolytic genes and operons in Bacillus subtilis: evidence for the presence of multiple levels of control of the gapA operon. Mol Microbiol 41:409–422[CrossRef]
LuesinkE. J., van HerpenR. E., GrossiordB. P., KuipersO. P., de VosW. M.1998; Transcriptional activation of the glycolytic las operon and catabolite repression of the gal operon in Lactococcus lactis are mediated by the catabolite control protein CcpA. Mol Microbiol 30:789–798[CrossRef]
MahrK., EstebanC. D., HillenW., TitgemeyerF., Pérez-MartínezG.2002; Cross communication between components of carbon catabolite repression of Lactobacillus casei and Bacillus megaterium. J Mol Microbiol Biotechnol 4:489–494
MonederoV., GosalbesM. J., Pérez-MartínezG.1997; Catabolite repression in Lactobacillus casei ATCC 393 is mediated by CcpA. J Bacteriol 179:6657–6664
MorenoM. S., SchneiderB. L., MaileR. R., WeylerW., SaierM. H., Jr. 2001; Catabolite repression mediated by the CcpA protein in Bacillus subtilis: novel modes of regulation revealed by whole-genome analyses. Mol Microbiol 39:1366–1381[CrossRef]
OlahG. A., TrakhanovS., TrewhellaJ., QuiochoF. A.1993; Leucine/isoleucine/valine-binding protein contracts upon binding of ligand. J Biol Chem 268:16241–16247
Pérez-MartínezG., KokJ., VenemaG., Van DijlJ. M., SmithH., BronS.1992; Protein export elements from Lactococcus lactis. Mol Gen Genet 234:401–411[CrossRef]
PosnoM., HeulvelmansP. T. H. M., LokmanB. C., PowelsP. H., LeerR. J., van LuijkN., van GienzenM. J. F.1991; Incompatibility of Lactobacillus vectors with replicons derived from small cryptic Lactobacillus plasmids and segregational instability of the induced vectors. Appl Environ Microbiol 57:1822–1828
SambrookJ., FritschE. F., ManiatisT.1989Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
SchumakerM. A., ChoiK. Y., ZalkinH., BrennanR. G.1994; Crystal structure of LacI member, PurR, bound to DNA: minor groove binding by α helices. Science 266:763–770[CrossRef]
SharffA. J., RodsethL. E., SpurlinoJ. C., QuiochoF. A.1992; Crystallographic evidence of a large ligand-induced hinge-twist motion between the two domains of the maltodextrin binding protein involved in active transport and chemotaxis. Biochemistry 31:10657–10663[CrossRef]
StanleyN. R., BrittonR. A., GrossmanA. D., LazazzeraB. A.2003; Identification of catabolite repression as a physiological regulator of biofilm formation by Bacillus subtilis by use of DNA microarrays. J Bacteriol 185:1951–1957[CrossRef]
VianaR., MonederoV., DossonnetV., VadeboncoeurC., Pérez-MartínezG., DeutscherJ.2000; Enzyme I and HPr from Lactobacillus casei: their role in sugar transport, carbon catabolite repression and inducer exclusion. Mol Microbiol 36:570–584
WenZ. T., BurneR. A.2002; Functional genomics approach to identifying genes required for biofilm development by Streptococcus mutans. Appl Environ Microbiol 68:1196–1203[CrossRef]