Complete Sequence of Plasmid pLH1 from Lactobacillus helveticus ATCC15009: Analysis Reveals the Presence of Regions Homologous to Other Native Plasmids from the Host Strain

Thompson, J. K.; Foley, Sophie; McConville, K.J.; Nicholson, Christine; Collins, Martin A.; Pridmore, R. David

doi:10.1006/plas.1999.1428

Cited by 19 publications

(16 citation statements)

References 32 publications

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“…The repA gene encodes a 12-kDa protein that shares homology with the C-terminal part of the replication proteins from the Lactobacillus sakei plasmid pSAK1, the Lactococcus lactis plasmid pCI2000, and the L. helveticus plasmids pLJ1 and pLH1 (32,54,56). Compared to those replication proteins, RepA lacks approximately 260 amino acids at its N terminus, the region that has strongest conservation and includes the helix-turn-helix motif that is considered to be involved in DNA binding (56). The repB gene encodes a 30-kDa protein and is homologous to proposed plasmid copy control proteins from the Enterococcus faecalis plasmids pAD1 and pAM373 and the Bacillus thuringiensis plasmid pAW63 (14,60,61).…”

Section: Resultsmentioning

confidence: 99%

Functional Analysis of Three Plasmids from Lactobacillus plantarum

Kranenburg

Golić

Bongers

et al. 2005

Appl Environ Microbiol

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Lactobacillus plantarum WCFS1 harbors three plasmids, pWCFS101, pWCFS102, and pWCFS103, with sizes of 1,917, 2,365, and 36,069 bp, respectively. The two smaller plasmids are of unknown function and contain replication genes that are likely to function via the rolling-circle replication mechanism. The host range of the pWCFS101 replicon includes Lactobacillus species and Lactococcus lactis, while that of the pWCFS102 replicon also includes Carnobacterium maltaromaticum and Bacillus subtilis. The larger plasmid is predicted to replicate via the theta-type mechanism. The host range of its replicon seems restricted to L. plantarum. Cloning vectors were constructed based on the replicons of all three plasmids. Plasmid pWCFS103 was demonstrated to be a conjugative plasmid, as it could be transferred to L. plantarum NC8. It confers arsenate and arsenite resistance, which can be used as selective markers.Lactic acid bacteria are used for the preservation of food and feed raw materials like milk, meat, and vegetables or other plant materials. Certain strains of lactic acid bacteria, in particular, strains from the genus Lactobacillus, have been attributed probiotic activities in humans and animals (30). Several lactic acid bacteria, including lactococci, streptococci, lactobacilli, and pediococci, are known to harbor plasmids. These may encode important traits like resistance to phages or antibiotics, lactose catabolism, and production of proteolytic enzymes or bacteriocins. Lactobacillus plantarum species often harbor several plasmids (49). Several of these have been sequenced (6,11,12,20,31,40,52,58). Although most of them are of unknown function, one plasmid encoding phage resistance (pMD5057, 10,877 bp) and another plasmid (pLKS, 2,025 bp) probably introduced from another source and coding for tetracycline resistance have been described previously (12,20). All are smaller than 11 kb and are predicted to replicate via the rolling-circle replication mechanism, except for the largest plasmid, pMD5057, which is predicted to replicate via the theta mechanism (12).The capacity for conjugal transfer is an important characteristic for plasmids. Self-transmissible conjugative plasmids have the ability to form effective cell-to-cell contact, while mobilizable plasmids are only able to prepare their DNA for transfer (38). Mobilization involves the action of a specific DNA-protein structure called the relaxosome to produce single-stranded cleavage at the nicking site (nic) within the origin of transfer (oriT) of the plasmid (38). To date, there is very little information on conjugation in lactobacilli. Sasaki et al. demonstrated conjugational transfer of the promiscuous thetareplicating plasmid pAM␤1 from Streptococcus faecalis to L. plantarum (51). Ahn et al. described an 8.5-kb chloramphenicol resistance plasmid which had been comobilized with pAM␤1 from L. plantarum to Carnobacterium maltaromaticum (3), previously known as Carnobacterium piscicola (43). To our knowledge, no conjugative L. plantarum plasmids have been reported...

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Section: Resultsmentioning

confidence: 99%

Functional Analysis of Three Plasmids from Lactobacillus plantarum

Kranenburg

Golić

Bongers

et al. 2005

Appl Environ Microbiol

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“…1; due to the high degree of identity (89%) between the pI9789::Tn552 and pSX267 replication regions, for clarity, only the longer pI9789::Tn552 sequence is shown. The replication initiation genes of these staphylococcal plasmids encode products similar (17 Յ Z Յ 43) to those of plasmids from other gram-positive genera, including pLS32 from Bacillus natto (52), pLJ1 (51), pLH1 (54), and pSAK1 (GenBank entry Z50862) from Lactobacillus species, and the enterococcal plasmids pAD1 (57), pCF10 (44), and pPD1 (14) ( Fig. 2A), a number of which have been proposed to utilize the theta mode of replication (see also reference 21).…”

Section: Resultsmentioning

confidence: 99%

Replication of Staphylococcal Multiresistance Plasmids

et al. 2000

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Based on structural and functional properties, three groups of large staphylococcal multiresistance plasmids have been recognized, viz., the pSK1 family, pSK41-like conjugative plasmids, and ␤-lactamase-heavy-metal resistance plasmids. Here we describe an analysis of the replication functions of a representative of each of these plasmid groups. The replication initiation genes from the Staphylococcus aureus plasmids pSK1, pSK41, and pI9789::Tn552 were found to be related to each other and to the Staphylococcus xylosus plasmid pSX267 and are also related to rep genes of several plasmids from other gram-positive genera. Nucleotide sequence similarity between pSK1 and pI9789::Tn552 extended beyond their rep genes, encompassing upstream divergently transcribed genes, orf245 and orf256, respectively. Our analyses revealed that genes encoding proteins related to the deduced orf245 product are variously represented, in several types of organization, on plasmids possessing six seemingly evolutionarily distinct types of replication initiation genes and including both thetamode and rolling-circle replicons. Construction of minireplicons and subsequent functional analysis demonstrated that orf245 is required for the segregational stability of the pSK1 replicon. In contrast, no gene equivalent to orf245 is evident on the conjugative plasmid pSK41, and a minireplicon encoding only the pSK41 rep gene was found to exhibit a segregational stability approaching that of the parent plasmid. Significantly, the results described establish that many of the large multiresistance plasmids that have been identified in clinical staphylococci, which were formerly presumed to be unrelated, actually utilize an evolutionarily related thetamode replication system.Clinical Staphylococcus aureus strains often harbor multiple plasmids, ranging from small rolling-circle (RC) replicating plasmids that are cryptic or encode only a single resistance determinant to larger multiresistance and conjugative plasmids (12,32,36). Three groups of multiresistance plasmids have been recognized in staphylococci. Isolates from the 1960s and 1970s were commonly found to carry multiresistance plasmids conferring resistance to penicillin and heavy metals or other inorganic ions (48). Such ␤-lactamase-heavy-metal resistance plasmids characteristically contain the ␤-lactamase-encoding transposon Tn552 or a derivative and operons mediating resistance to arsenical, cadmium, and/or mercuric ions (36). Some ␤-lactamase-heavy-metal resistance plasmids also contain Tn551, conferring resistance to macrolide-lincosamide-streptogramin type B antibiotics (33); an IS256-bounded composite aminoglycoside resistance transposon, Tn4001 (30); and/or a qacA or qacB antiseptic and disinfectant multidrug resistance determinant (30).pSK41-like conjugative multiresistance plasmids were first detected in strains isolated in the mid 1970s. Such plasmids commonly contain a Tn4001 hybrid structure (6) and IS257-flanked cointegrated copies of small plasmids, such as the aminoglycoside resist...

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“…Like other plasmids (14,49), pRV500 appears to be a composite structure containing DNA segments from different sources. Some ORFs are shared by plasmids from both subfamilies pUCL22 and pUCL287; this applies to orf6 in pRV500 and ORFs of unknown function around the gene for a sitespecific recombinase (orf10).…”

Section: Discussionmentioning

confidence: 99%

“…A conserved feature is a palindromic sequence of about 40 bp, located downstream of orf13 in pRV500; it is identical at more than 90% where indicated in plasmids (Fig. 4) and could serve as a terminator for the upstream ORFs, as in pLH1 (49).…”

Section: Vol 69 2003 a Theta-type Plasmid From Lactobacillus Sakei mentioning

confidence: 99%

“…Both RC and theta types of replicons have been reported in lactobacilli. In L. sakei, RC plasmids closely related to those of Lactobacillus curvatus (31) and Lactobacillus plantarum (9) have been identified, and the partial sequence of a potential theta-replicating plasmid is known, namely, pSAK1 (GenBank Z50862), belonging to the pLS32 family (49). In this paper, we present the analysis of the complete sequence of a new L. sakei plasmid.…”

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Characterization of a Theta-Type Plasmid from Lactobacillus sakei : a Potential Basis for Low-Copy-Number Vectors in Lactobacilli

Alpert¹,

Coq²,

Malleret³

et al. 2003

Appl Environ Microbiol

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The complete nucleotide sequence of the 13-kb plasmid pRV500, isolated from Lactobacillus sakei RV332, was determined. Sequence analysis enabled the identification of genes coding for a putative type I restrictionmodification system, two genes coding for putative recombinases of the integrase family, and a region likely involved in replication. The structural features of this region, comprising a putative ori segment containing 11-and 22-bp repeats and a repA gene coding for a putative initiator protein, indicated that pRV500 belongs to the pUCL287 subfamily of theta-type replicons. A 3.7-kb fragment encompassing this region was fused to an Escherichia coli replicon to produce the shuttle vector pRV566 and was observed to be functional in L. sakei for plasmid replication. The L. sakei replicon alone could not support replication in E. coli. Plasmid pRV500 and its derivative pRV566 were determined to be at very low copy numbers in L. sakei. pRV566 was maintained at a reasonable rate over 20 generations in several lactobacilli, such as Lactobacillus curvatus, Lactobacillus casei, and Lactobacillus plantarum, in addition to L. sakei, making it an interesting basis for developing vectors. Sequence relationships with other plasmids are described and discussed.Lactobacilli belong to the lactic acid bacteria (LAB) group. The genus comprises several species important for the food industry, among which is Lactobacillus sakei, one of the most important bacterial species involved in meat fermentation and preservation. L. sakei belongs to the dominant microflora naturally occurring on vacuum-packed meat and is also used in Western Europe as starter cultures for fermented products (especially sausage) because of its ability to rapidly ferment carbohydrates in the meat environment (11,26).Most lactobacilli harbor one or more plasmids, the size of which can vary from 1.2 to 150 kb (for a review, see reference 50). Different functions have already been found on these plasmids, among which are genes for lactose metabolism, bacteriocin synthesis, exopolysaccharide production, or DNA restriction-modification (R-M). With the development of molecular biology and functional genomics of lactobacilli, interest is growing for the characterization of replicons themselves as potential useful (food-grade) vectors. Two modes of DNA replication are used by circular bacterial plasmids, namely, rolling circle (RC) and theta. RC plasmids have been assumed to be the most widespread in gram-positive bacteria. However, recent years have seen the characterization of a large number of theta-replicating plasmids from gram-positive, and especially from LAB, hosts. Several mechanisms are known for the initiation of theta replication. Theta replicons can be classified according to their dependence on three key components. These are plasmid-encoded initiator Rep proteins necessary for strand opening and/or interaction with other components of the initiation complex, origins of replication (generically termed ori) with specific DNA structural organization...

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Complete Sequence of Plasmid pLH1 from Lactobacillus helveticus ATCC15009: Analysis Reveals the Presence of Regions Homologous to Other Native Plasmids from the Host Strain

Cited by 19 publications

References 32 publications

Functional Analysis of Three Plasmids from Lactobacillus plantarum

Functional Analysis of Three Plasmids from Lactobacillus plantarum

Replication of Staphylococcal Multiresistance Plasmids

Characterization of a Theta-Type Plasmid from Lactobacillus sakei : a Potential Basis for Low-Copy-Number Vectors in Lactobacilli

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