Double-glycine-type leader peptides direct secretion of bacteriocins by ABC transporters: colicin V secretion in Lactococcus lactis.
Many non-lantibiotic bacteriocins of lactic acid bacteria are produced as precursors which have N-terminal leader peptides that share similarities in amino acid sequence and contain a conserved processing site of two glycine residues in positions -1 and -2. A dedicated ATP-binding cassette (ABC) transporter is responsible for the proteolytic cleavage of the leader peptides and subsequent translocation of the bacteriocins across the cytoplasmic membrane. To investigate the role that these leader peptides play in the recognition of the precursor by the ABC transporters, the leader peptides of leucocin A, lactococcin A or colicin V were fused to divergicin A, a bacteriocin from Carnobacterium divergens that is secreted via the cell's general secretion pathway. Production of divergicin was monitored when these fusion constructs were introduced into Leuconostoc gelidum, Lactococcus lactis and Escherichia coli, which carry the secretion apparatus for leucocin A, lactococcins A and B, and colicin V, respectively. The different leader peptides directed the production of divergicin in the homologous hosts. In some cases production of divergicin was also observed when the leader peptides were used in heterologous hosts. For ABC-transporter-dependent secretion in E. coli the outer membrane protein TolC was required. Using this strategy, colicin V was produced in L. lactis by fusing this bacteriocin behind the leader peptide of leucocin A.
van Belkum
M J
MJ
Department of Agricultural, Food and Nutritional Science, University of Alberta, Edmonton, Canada.
Worobo
R W
RW
Stiles
M E
ME
D018528:ATP-Binding Cassette Transporters / Q000235:genetics* / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000502:physiology; D001430:Bacteriocins / Q000378:metabolism*; D001692:Biological Transport; D003087:Colicins / Q000378:metabolism / Q000494:pharmacology; D029968:Escherichia coli Proteins; D005998:Glycine / Q000378:metabolism; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D007934:Leuconostoc / Q000378:metabolism; D026901:Membrane Transport Proteins; D021382:Protein Sorting Signals / Q000737:chemistry / Q000502:physiology*
D018528:ATP-Binding Cassette Transporters; D001425:Bacterial Outer Membrane Proteins; D001430:Bacteriocins; D003087:Colicins; D029968:Escherichia coli Proteins; D026901:Membrane Transport Proteins; D021382:Protein Sorting Signals; C093812:dvnA protein, Carnobacterium divergens; C073126:tolC protein, E coli; C069325:lactococcin A; D005998:Glycine