The lactococcin G immunity protein recognizes specific regions in both peptides constituting the two-peptide bacteriocin lactococcin G.
Lactococcin G and enterocin 1071 are two homologous two-peptide bacteriocins. Expression vectors containing the gene encoding the putative lactococcin G immunity protein (lagC) or the gene encoding the enterocin 1071 immunity protein (entI) were constructed and introduced into strains sensitive to one or both of the bacteriocins. Strains that were sensitive to lactococcin G became immune to lactococcin G when expressing the putative lactococcin G immunity protein, indicating that the lagC gene in fact encodes a protein involved in lactococcin G immunity. To determine which peptide or parts of the peptide(s) of each bacteriocin that are recognized by the cognate immunity protein, combinations of wild-type peptides and hybrid peptides from the two bacteriocins were assayed against strains expressing either of the two immunity proteins. The lactococcin G immunity protein rendered the enterococcus strain but not the lactococcus strains resistant to enterocin 1071, indicating that the functionality of the immunity protein depends on a cellular component. Moreover, regions important for recognition by the immunity protein were identified in both peptides (Lcn-alpha and Lcn-beta) constituting lactococcin G. These regions include the N-terminal end of Lcn-alpha (residues 1 to 13) and the C-terminal part of Lcn-beta (residues 14 to 24). According to a previously proposed structural model of lactococcin G, these regions will be positioned adjacent to each other in the transmembrane helix-helix structure, and the model thus accommodates the present results.
Oppegård
Camilla
C
Department of Molecular Biosciences, University of Oslo, P.O. Box 1041 Blindern, 0316 Oslo, Norway. camilla.oppegard@imbv.uio.no
Emanuelsen
Linda
L
Thorbek
Lisbeth
L
Fimland
Gunnar
G
Nissen-Meyer
Jon
J
D000595:Amino Acid Sequence; D001430:Bacteriocins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D013293:Enterococcus faecalis / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D017045:Lactococcus / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D013294:Lactococcus lactis / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics; D017386:Sequence Homology, Amino Acid
D001430:Bacteriocins; D017931:DNA Primers; D004269:DNA, Bacterial; C407407:enterocin 1071A; C407408:enterocin 1071B; C110915:lactococcin G