Inducible bacteriocin production in Lactobacillus is regulated by differential expression of the pln operons and by two antagonizing response regulators, the activity of which is enhanced upon phosphorylation.
Expression of the five (pln) operons involved in the bacteriocin production of Lactobacillus plantarum C11 is regulated by a so-called pheromone-based signal-transducing network, in which the peptide pheromone (PlnA) induces bacteriocin production through the action of a histidine protein kinase (PlnB) and two antagonizing response regulators (PlnC as an activator and PlnD as a negative regulator). All pln-regulated promoters contain a conserved pair of direct repeats that serve as binding sites for PlnC and PlnD. In the present work, we show that the five PlnA-responsive operons are differentially expressed with regard to both timing and strength, and that the pheromone triggers a strong autoactivating loop of the regulatory unit (plnABCD) during an early stage of induction that gradually leads to enhanced activation of the other operons. The transport operon (plnGHSTUV), which is involved in the secretion of the pheromone and bacteriocins, is also expressed relatively early upon induction, but is quickly turned off soon after peak expression. Further investigation of the various promoters revealed that, although subtle differences within the promoter regions could account for the observed differential regulation, the presence of a downstream promoter-proximal sequence in one promoter was found to cause delayed peak activity. How phosphorylation regulates the activity of the pln response regulators was also accessed by direct mutagenesis at their phosphorylation sites. It was found that the two response regulators exert activity at two different levels: a low level when they are not phosphorylated and an elevated level when they are phosphorylated. The present data demonstrate that bacteriocin production in L. plantarum C11 is a highly regulated process, in which different regulatory mechanisms are applied to fine tune the timing and strength of expression of the five pln operons.
Diep
Dzung B
DB
Laboratory of Microbial Gene Technology, Department of Chemistry, Agricultural University of Norway, Norway.
Myhre
Ronny
R
Johnsborg
Ola
O
Aakra
Agot
A
Nes
Ingolf F
IF
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001430:Bacteriocins / Q000096:biosynthesis*; D015964:Gene Expression Regulation, Bacterial; D007778:Lactobacillus / Q000235:genetics / Q000254:growth & development* / Q000378:metabolism; D008969:Molecular Sequence Data; D009876:Operon; D010675:Pheromones / Q000494:pharmacology*; D010766:Phosphorylation; D011401:Promoter Regions, Genetic; D015398:Signal Transduction; D014158:Transcription, Genetic
D001426:Bacterial Proteins; D001430:Bacteriocins; D010675:Pheromones