Use of two-dimensional electrophoresis To study differential protein expression in divercin V41-resistant and wild-type strains of Listeria monocytogenes.
The use of bacteriocins from food-grade lactic acid bacteria to fight against the food-borne pathogen Listeria monocytogenes has been gaining interest. However, the emergence of resistant cells is frequently reported when Listeria is exposed to such antibacterials. A two-dimensional electrophoresis study of whole-cell protein expression of Listeria monocytogenes variants sensitive or resistant to the action of a bacteriocin produced by Carnobacterium divergens V41, divercin V41, is reported in this paper. The resistant variant obtained from the sensitive strain of L. monocytogenes P was also resistant to piscicocins V1 and SF668, but remained sensitive to nisin. Its growth rate was 50% less than the sensitive strain, and the MIC for it was 10(4) times higher. No reversion of the resistance was observed after 20 successive cultures in the absence of divercin V41. Comparison of the protein patterns by two-dimensional gel electrophoresis analysis showed clear differences. In the resistant variant pattern, at least nine spots had disappeared and eight new ones were observed. One of the newly synthesized proteins was identified as a flagellin of L. monocytogenes. Direct interaction between flagellin and divercin V41 was not evidenced. Intracellular synthesis of flagellin is probably an indirect effect of a modification in transcriptional regulation with widespread effects through a sigma factor. An intense protein, only present in the sensitive strain, was identified as a non-heme iron-binding ferritin displaying strong similarities to Dps proteins. Common modifications in the transcriptional regulation for these two proteins are discussed.
Duffes
F
F
Laboratoire de Recherches de Technologie Laitière, Institut National de la Recherche Agronomique, 35042 Rennes Cedex, France.
Jenoe
P
P
Boyaval
P
P
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001426:Bacterial Proteins / Q000032:analysis* / Q000737:chemistry / Q000494:pharmacology; D001430:Bacteriocins / Q000494:pharmacology*; D004352:Drug Resistance, Microbial / Q000235:genetics; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005407:Flagella / Q000737:chemistry / Q000648:ultrastructure; D014644:Genetic Variation; D007777:Lactobacillaceae; D008089:Listeria monocytogenes / Q000145:classification / Q000187:drug effects* / Q000235:genetics*; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D012703:Serotyping
D000900:Anti-Bacterial Agents; D001426:Bacterial Proteins; D001430:Bacteriocins; C116372:divercin V41