The cellular location and effect on nisin immunity of the NisI protein from Lactococcus lactis N8 expressed in Escherichia coli and L. lactis

Qiao, Mingqiang; Immonen, Tiina; Koponen, Olli; Saris, Per E. J.

doi:10.1016/0378-1097(95)00238-z

Cited by 42 publications

(42 citation statements)

References 18 publications

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“…It is conceivable that the putative lipoprotein encoded by pspQ also functions to mediate resistance in the producing strains by sequestering the lantibiotic, preventing interaction with lipid II. A similar function has been proposed for other lipoproteins encoded in lantibiotic biosynthetic gene clusters (37,38).…”

Section: Manipulating the Regulatory Genes Of The Psp Gene Cluster Resupporting

confidence: 61%

The antibiotic planosporicin coordinates its own production in the actinomycete Planomonospora alba

Sherwood

Bibb

2013

Proc. Natl. Acad. Sci. U.S.A.

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Planosporicin is a ribosomally synthesized, posttranslationally modified peptide lantibiotic produced by the actinomycete Planomonospora alba. It contains one methyl-lanthionine and four lanthionine bridges and inhibits cell wall biosynthesis in other Gram-positive bacteria probably by binding to lipid II, the immediate precursor for cell wall biosynthesis. Planosporicin production, which is encoded by a cluster of 15 genes, is confined to stationary phase in liquid culture and to the onset of morphological differentiation when P. alba is grown on agar. This growth phase-dependent gene expression is controlled transcriptionally by three pathway-specific regulatory proteins: an extracytoplasmic function σ factor (PspX), its cognate anti-σ factor (PspW), and a transcriptional activator (PspR) with a C-terminal helix-turn-helix DNA-binding domain. Using mutational analysis, S1 nuclease mapping, quantitative RT-PCR, and transcriptional fusions, we have determined the direct regulatory dependencies within the planosporicin gene cluster and present a model in which subinhibitory concentrations of the lantibiotic function in a feed-forward mechanism to elicit high levels of planosporicin production. We show that in addition to acting as an antibiotic, planosporicin can function as an extracellular signaling molecule to elicit precocious production of the lantibiotic, presumably ensuring synchronous and concerted lantibiotic biosynthesis in the wider population and, thus, the production of ecologically effective concentrations of the antibiotic.

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Section: Manipulating the Regulatory Genes Of The Psp Gene Cluster Resupporting

confidence: 61%

The antibiotic planosporicin coordinates its own production in the actinomycete Planomonospora alba

Sherwood

Bibb

2013

Proc. Natl. Acad. Sci. U.S.A.

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“…The B. subtilis nisin tolerance level provided by the nisin immunity genes suggests the additive action of two independent systems, the lipoprotein NisI and the transporter NisFEG. Nevertheless, we cannot exclude the effect of additional factors that contribute to nisin immunity in the nisin producer L. lactis, for which a rather cooperative effect of NisI and NisFEG was discussed (35,50). Remarkably, the transfer of all components of the nisin immunity system to a subtilin-producing B. subtilis host was successful.…”

Section: Discussionmentioning

confidence: 95%

Function of Lactococcus lactis Nisin Immunity Genes nisI and nisFEG after Coordinated Expression in the Surrogate Host Bacillus subtilis

Stein

Heinzmann

Solovieva

et al. 2003

Journal of Biological Chemistry

162

240

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Nisin-producing Lactococcus lactis strains show a high degree of resistance to the action of nisin, which is based upon expression of the self-protection (immunity) genes nisI, nisF, nisE, and nisG. Different combinations of nisin immunity genes were integrated into the chromosome of a nisin-sensitive Bacillus subtilis host strain under the control of an inducible promoter. For the recipient strain, the highest level of acquired nisin tolerance was achieved after coordinated expression of all four nisin immunity genes. But either the lipoprotein NisI or the ABC transporter-homologous system Nis-FEG, respectively, were also able to protect the Bacillus host cells. The acquired immunity was specific to nisin and provided no tolerance to subtilin, a closely related lantibiotic. Quantitative in vivo peptide release assays demonstrated that NisFEG diminished the quantity of cell-associated nisin, providing evidence that one role of NisFEG is to transport nisin from the membrane into the extracellular space. NisI solubilized from B. subtilis membrane vesicles and recombinant hexahistidinetagged NisI from Escherichia coli interacted specifically with nisin and not with subtilin. This suggests a function of NisI as a nisin-intercepting protein.In recent years peptide antibiotics have gained increasing attention as therapeutics (1, 2) and food preservatives (3). Nisin represents the most prominent member of lantibiotics, peptide antibiotics with intramolecular lanthionine bridges (4 -9). The nisin producer Lactococcus lactis 6F3 contains a gene cluster encoding proteins for the biosynthesis and transport (10 -14), immunity (15), and regulation (16 -18) of nisin. Subtilin (19,20) and ericin S (21) produced by Bacillus subtilis ATCC 6633 and A1/3, respectively, are closely related lantibiotics. Lantibiotics form voltage-dependent pores in the bacterial cytoplasmic membrane that are lethal for the target cells but also for the producer. For nisin, the mode of action was investigated in several model systems such as black lipid bilayers and membrane vesicles (22)(23)(24)(25). Recent findings demonstrated that specific binding of nisin to the cell wall precursor lipid II coincides with pore formation (26, 27). Specific selfprotection (immunity) mechanisms are necessary to protect the lantibiotic-producing organisms from the action of their own lantibiotics. For nisin and subtilin (28) Although numerous genes involved in lantibiotic immunity are known, the mechanism by which the encoded proteins confer immunity remain unclear. For full nisin or subtilin immunity, both are required, i.e. the lipoprotein as well as the immunity transporter. The lack of each component diminished the tolerance to nisin (35) or subtilin (28) significantly. Here we report for the first time on the establishment of nisin immunity in the heterologous host B. subtilis. Functional analyses of its different components provided evidence that NisI acts as a nisin-sequestering protein and that NisFEG acts as a nisin exporter that expels nisin molecules fro...

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“…It is known that all lanthionines are needed for nisin activity and if the leader is not cleaved by NisP, the fully modified nisin precursor still containing the N-terminal leader is inactive (van der Meer et al, 1993 ;Qiao et al, 1995). Interestingly, the His-tag seemed to impair the inductive capacity of nisin as no active His-tagged nisin could be produced unless cells were induced with intact nisin.…”

Section: Discussionmentioning

confidence: 99%

NisB is required for the dehydration and NisC for the lanthionine formation in the post-translational modification of nisin

et al. 2002

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Nisin produced by Lactococcus lactis subsp. lactis is a 34-residue antibacterial polypeptide and belongs to a group of post-translationally modified peptides, lantibiotics, with dehydrated residues and cyclic amino acids, lanthionines. These modifications are supposed to be made by enzymes encoded by lanB and lanC genes, found only in biosynthetic operons encoding lantibiotics. To analyse the extent of modification, His-tagged nisin precursors were expressed in nisB and nisC mutant strains. The His-tagged nisin precursors were purified from the cytoplasm of the cells, as lack of NisB or NisC activity impaired translocation of the nisin precursor. The purified His-tagged polypeptides were analysed with trypsin digestion followed by nisin bioassay, SDS-PAGE, Nterminal sequencing and mass spectroscopy. According to the results, nisin precursors from the strain lacking NisB activity were totally unmodified, whereas nisin precursors from the strain lacking NisC activity, but having NisB activity, were dehydrated and devoid of normal lanthionine formation. This is the first experimental evidence showing that NisB is required for dehydration and NisC for correct lanthionine formation in nisin maturation.

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The cellular location and effect on nisin immunity of the NisI protein from Lactococcus lactis N8 expressed in Escherichia coli and L. lactis

Cited by 42 publications

References 18 publications

The antibiotic planosporicin coordinates its own production in the actinomycete Planomonospora alba

The antibiotic planosporicin coordinates its own production in the actinomycete Planomonospora alba

Function of Lactococcus lactis Nisin Immunity Genes nisI and nisFEG after Coordinated Expression in the Surrogate Host Bacillus subtilis

NisB is required for the dehydration and NisC for the lanthionine formation in the post-translational modification of nisin

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