The complete nucleotide sequence of the 3475 bp plasmid pCD3.4 from Carnobacterium divergens LV13, which encodes the bacteriocin divergicin A, was determined. Nucleotide sequence, deletion and complementation analyses revealed the presence of a trans-acting replication protein, RepA, and DNA sequences involved in plasmid replication and copy-number control. The DNA region preceding the repA gene probably contains the origin of replication. This sequence includes four and a half direct repeats (iterons) of 22 bp, to which RepA is thought to bind, and an AT-rich region containing a 12 bp repeat, at which initiation of DNA might occur. Further upstream of this sequence resides a fifth iteron required for optimal plasmid replication. The RepA protein shows homology to replication proteins of the pUCL287 subfamily of theta-type replicons. Two ORFs were found downstream of the repA gene that could be deleted without affecting replication and stability of the plasmid. pCD3.4 has a narrow host range, and could only be maintained in Carnobacterium spp.; however, a mutant of the plasmid was obtained that enabled the pCD3.4 replicon to replicate in Enterococcus faecium, but not in Carnobacterium spp. The mutation was located in the C-terminal region of the RepA protein, changing a proline into a serine. This is believed to be the first example of such plasmid-host-range modulation in Gram-positive bacteria.
AbhyankarM. M, ReddyJ. M, SharmaR, BüllesbachE,
BastiaD. 2004; Biochemical investigations of control of replication initiation of plasmid R6K. J Biol Chem 279:6711–6719
AhnC, StilesM. E.
1990; Plasmid-associated bacteriocin production by a strain of Carnobacterium piscicola from meat. Appl Environ Microbiol 56:2503–2510
AubertS, DykeK. G, SolhN. E.
1998; Analysis of two Staphylococcus epidermidis plasmids coding for resistance to streptogramin A. Plasmid 40:238–242[CrossRef]
BenachourA, FrèreJ,
NovelG. 1995; pUCL287 plasmid from Tetragenococcus halophilus (Pediococcus halophilus) ATCC 33315 represents a new theta-type replicon family of lactic acid bacteria. FEMS Microbiol Lett 128:167–176[CrossRef]
BenachourA, NovelG, AuffrayY,
FrèreJ,
FlahautS. 1997; Molecular analysis of the replication region of the theta-replicating plasmid pUCL287 from Tetragenococcus (Pediococcus) halophilus ATCC 33315. Mol Gen Genet 255:504–513[CrossRef]
BruandC, Le ChatelierE, EhrlichS. D, JannièreL. 1993; A fourth class of theta-replicating plasmids: the pAM β 1 family from Gram-positive bacteria. Proc Natl Acad Sci U S A 90:11668–11672[CrossRef]
ChurchwardG, LinderP, CaroL.
1983; The nucleotide sequence of replication and maintenance functions encoded by plasmid pSC101. Nucleic Acids Res 11:5645–5659[CrossRef]
DattaH. J, KhatriG. S, BastiaD.
1999; Mechanism of recruitment of DnaB helicase to the replication origin of the plasmid pSC101. Proc Natl Acad Sci U S A 96:73–78[CrossRef]
Fernández-TresguerresM. E,
MartínM,
García de ViedmaD,
GiraldoR, Díaz-OrejasR. 1995; Host growth temperature and a conservative amino acid substitution in the replication protein of pPS10 influence plasmid host range. J Bacteriol 177:4377–4384
FoleyS, BronS, VenemaG, DalyC, FitzgeraldG. F.
1996; Molecular analysis of the replication origin of the Lactococcus lactis plasmid pCI305. Plasmid 36:125–141[CrossRef]
FranzC. M. A. P, WoroboR. W, QuadriL. E. N, SchillingerU, HolzapfelW. H, VederasJ. C, StilesM. E.
1999; Atypical genetic locus associated with constitutive production of enterocin B by Enterococcus faecium BFE900. Appl Environ Microbiol 65:2170–2178
FujimotoS, TomitaH, WakamatsuE, TanimotoK, IkeY.
1995; Physical mapping of the conjugative bacteriocin plasmid pPD1 of Enterococcus faecalis and identification of the determinant related to the pheromone response. J Bacteriol 177:5574–5581
GassonM. J.
1983; Plasmid complements of Streptococcus lactis NCDO712 and other lactic streptococci after protoplast-inducing curing. J Bacteriol 154:1–9
HayesF, VosP, FitzgeraldG. F, DalyC,
de VosW. M. 1991; Molecular organization of the minimal replicon of novel, narrow-host-range, lactococcal plasmid pCI305. Plasmid 25:16–26[CrossRef]
HedbergP. J, LeonardB. A, RuhfelR. E, DunnyG. M.
1996; Identification and characterization of the genes of Enterococcus faecalis plasmid pCF10 involved in replication and in negative control of pheromone-inducible conjugation. Plasmid 35:46–57[CrossRef]
HoloH, NesI. F.
1989; High-frequency transformation, by electroporation, of Lactococcus lactis subsp. cremoris grown in glycine in osmotically stabilized media. Appl Environ Microbiol 173:3879–3887
JiangY, PacekM, HelinskiD. R, KoniecznyI, ToukdarianA.
2003; A multifunctional plasmid-encoded replication initiation protein both recruits and positions active helicase at the replication origin. Proc Natl Acad Sci U S A 100:8692–8697[CrossRef]
KanataniK, TaharaT, OshimuraM, SanoK, UmezawaC.
1995; Identification of the replication region of Lactobacillus acidophilus plasmid pLA103. FEMS Microbiol Lett 133:127–130[CrossRef]
KearneyK, FitzgeraldG. F, SeegersJ. F. M. L.
2000; Identification and characterization of an active plasmid partition mechanism for the novel Lactococcus lactis plasmid pCI2000. J Bacteriol 182:30–37[CrossRef]
KiewietR, KokJ, SeegersJ. F. M. L, VenemaG, BronS.
1993b; The mode of replication is a major factor in segregational plasmid instability in Lactococcus lactis . Appl Environ Microbiol 59:358–364
KimY.-J, MeyerR. J.
1991; An essential iteron-binding protein required for plasmid R1162 replication induces localized melting within the origin at a specific site in AT-rich DNA. J Bacteriol 173:5539–5545
Le ChatelierE, EhrlichS. D, JannièreL. 1993; Biochemical and genetic analysis of the unidirectional theta replication of the S. agalactiae plasmid pIP501. Plasmid 29:50–56[CrossRef]
LeenhoutsK. J, TolnerB, BronS, KokJ, VenemaG, SeegersJ. F. M. L.
1991; Nucleotide sequence and characterization of the broad-host-range lactococcal plasmid pWVO1. Plasmid 26:55–66[CrossRef]
MaestroB, SanzJ. M, FaelenM, CouturierM, Díaz-OrejasR,
Fernández-TresguerresE. 2002; Modulation of pPS10 host range by DnaA. Mol Microbiol 46:223–234[CrossRef]
MaestroB, SanzJ. M, Díaz-OrejasR,
Fernández-TresguerresE. 2003; Modulation of pPS10 host range by plasmid-encoded RepA initiator protein. J Bacteriol 185:1367–1375[CrossRef]
MeijerW. J. J, VenemaG, BronS,
de BoerA. J,
van TongerenS. 1995; Characterization of the replication region of the Bacillus subtilis plasmid pLS20: a novel type of replicon. Nucleic Acids Res 23:3214–3223[CrossRef]
SørvigE, SkaugenM, NaterstadK, EijsinkV. G, AxelssonL.
2005; Plasmid p256 from Lactobacillus plantarum represents a new type of replicon in lactic acid bacteria, and contains a toxin–antitoxin-like plasmid maintenance system. Microbiology 151:421–431[CrossRef]
StudierF. W, MoffatB.
1986; Use of bacteriophage T7 RNA polymerase to direct selective high expression of cloned genes. J Mol Biol 189:113–130[CrossRef]
TaborS, RichardsonC. C.
1985; A bacteriophage T7 RNA polymerase promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A 82:1074–1078[CrossRef]
van BelkumM. J.,
StilesM. E.
1995; Molecular characterization of genes involved in the production of the bacteriocin leucocin A from Leuconostoc gelidum . Appl Environ Microbiol 61:3573–3579
van BelkumM. J,
WoroboR. W, StilesM. E.
1997; Double-glycine-type leader peptides direct secretion of bacteriocins by ABC transporters: colicin V secretion in Lactococcus lactis . Mol Microbiol 23:1293–1301[CrossRef]
van de GuchteM,
van der VossenJ. M. B. M,
KokJ, VenemaG.
1989; Construction of a lactococcal expression vector: expression of hen egg white lysozyme in Lactococcus lactis subsp. lactis . Appl Environ Microbiol 55:224–228
WeaverK. E, ClewellD. B, AnF.
1993; Identification, characterization, and nucleotide sequence of a region of Enterococcus faecalis pheromone-responsive plasmid pAD1 capable of autonomous replication. J Bacteriol 175:1900–1909
WoroboR. W, SailerM, RoyK. L, VederasJ. C, StilesM. E,
van BelkumM. J. 1995; A signal peptide secretion-dependent bacteriocin from Carnobacterium divergens . J Bacteriol 177:3143–3149