title: Glucose-1-phosphate utilization by Listeria monocytogenes is PrfA dependent and coordinately expressed with virulence factors.
abstract: Virulence genes of the facultative intracellular pathogen Listeria monocytogenes are coordinately regulated by the activator protein PrfA, encoded by prfA, a member of the cyclic AMP receptor protein family of bacterial transcription factors. We found that prfA* mutants that constitutively overexpress the virulence regulon due to a Gly145Ser substitution in PrfA (M.-T. Ripio, G. Domínguez-Bernal, M. Lara, M. Suárez, and J.-A. Vázquez-Boland, J. Bacteriol. 179:1533-1540, 1997) rapidly utilized glucose-1-phosphate (G-1-P) as a carbon source for growth, in contrast to wild-type strains, which characteristically do not. Wild-type strains acquired the capacity for readily metabolizing G-1-P upon exposure to environmental conditions that activate the expression of prfA and PrfA-dependent virulence genes (i.e., culture at 37 degrees C in charcoal-treated medium). In these strains, G-1-P utilization followed an expressional pattern identical to that of virulence genes controlled by PrfA, with repression at 20 degrees C. Tn917 insertions in L. monocytogenes mutants selected for G-1-P utilization deficiency mapped to the plcA-prfA operon, a deltaprfA strain was totally unable to utilize G-1-P, and trans complementation with prfA constructs restored the ability to efficiently metabolize and grow on G-1-P to these mutants. Thus, G-1-P utilization by L. monocytogenes is under the tight positive control of the central virulence regulator, PrfA, and is coexpressed with PrfA-dependent pathogenicity determinants. It was recently reported that readily utilized carbohydrates, such as glucose or cellobiose, repress virulence genes in L. monocytogenes. We confirmed this but, interestingly, found that G-1-P does not inhibit expression of the PrfA regulon, indicating that this sugar follows a catabolic pathway that bypasses the repressor mechanism triggered by other readily metabolized carbon sources. PrfA dependence and coexpression with virulence genes suggest that utilization of exogenous G-1-P may be relevant to Listeria pathogenesis. G-1-P is the precursor metabolite and primary degradation product of glycogen and is therefore available within the mammalian cell. Based on our results, we hypothesize that G-1-P could play an important role as a growth substrate for intracellular Listeria.
authors.affiliation: Unidad de Microbiología e Immunología, Facultad de Veterinaria, Universidad Complutense, Madrid, Spain.
authors.forename: M T
authors.initials: MT
authors.lastname: Ripio
authors.forename: K
authors.initials: K
authors.lastname: Brehm
authors.forename: M
authors.initials: M
authors.lastname: Lara
authors.forename: M
authors.initials: M
authors.lastname: Suárez
authors.forename: J A
authors.initials: JA
authors.lastname: Vázquez-Boland
mesh_terms: D019943:Amino Acid Substitution; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D002475:Cellobiose / Q000378:metabolism; D002874:Chromosome Mapping; D004251:DNA Transposable Elements; D015964:Gene Expression Regulation, Bacterial; D005816:Genetic Complementation Test; D005947:Glucose / Q000378:metabolism; D005958:Glucosephosphates / Q000378:metabolism*; D008089:Listeria monocytogenes / Q000235:genetics* / Q000378:metabolism* / Q000472:pathogenicity; D016254:Mutagenesis, Insertional; D010454:Peptide Termination Factors; D018085:Regulon; D015534:Trans-Activators / Q000235:genetics* / Q000378:metabolism*; D014774:Virulence / Q000235:genetics
chemical_list: D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D005958:Glucosephosphates; D010454:Peptide Termination Factors; D015534:Trans-Activators; D002475:Cellobiose; C031590:glucose-1-phosphate; D005947:Glucose