Nucleotide sequence and structural organization of the small, broad-host-range plasmid pCI411 from Leuconostoc lactis 533

Coffey, Aidan; Harrington, Aidan; Kearney, Karen; Daly, Charles; Fitzgerald, Gerald F.

doi:10.1099/13500872-140-9-2263

Cited by 42 publications

(21 citation statements)

References 23 publications

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“…since they have a region whose nucleotide sequence is very similar to that of well 464 characterized oriTs but lack a cognate putative relaxase gene; such is the case of 465 plasmids pA1 from Lactobacillus plantarum (Vujcic and Topisirovic, 1993) and 466 pCI411 from Leuconoctoc lactis (Coffey et al, 1994). This, in conjunction to the oriTs 467 of some RCR-plasmids being defined as site-specific plasmid recombination (Pre) 468 conditions.…”

Section: Recognition Of Heterologous Orits By Mobm Protein 420mentioning

confidence: 99%

Nicking activity of the pMV158 MobM relaxase on cognate and heterologous origins of transfer

Fernández-López

Lorenzo-Díaz

Pérez-Luque

et al. 2013

Plasmid

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The MobM relaxase (494 amino acids) encoded by the promiscuous streptococcal plasmid pMV158 recognizes the plasmid origin of transfer, oriTpMV158, and converts supercoiled pMV158 DNA into relaxed molecules by cleavage of the phosphodiester bond of a specific dinucleotide within the sequence 5'-GTGTG/TT-3' ("/" being the nick site). After cleavage, the protein remains stably bound to the 5´-end of the nick site. Band-shift assays with single-stranded oligonucleotides and sizeexclusion chromatography allowed us to show that MobM was able to generate specific complexes with one of the inverted repeats of the oriTpMV158, presumably extruded as stem-loop structure. A number of tests have been performed to attain a better characterization of the nicking activity of MobM and its linkage with its target DNA. The optimal pH for DNA relaxation was found to be 6.5. Upon nicking, gel retardation assays showed that MobM formed stable complexes with its target DNA. Moreover, MobM bound to relaxed pMV158 molecules were visualized by electron microscopy. The staphylococcal plasmids pUB110 and pE194, and the streptococcal plasmid pDL287 harbour putative oriTs and may encode Mob proteins homologous to MobM. The oriTpUB110, oriTpDL287, and oriTpE194 sequences share 100%, 70%, and 67% (in a 43-nucleotide stretch and allowing a 3-bp gap) identity to oriTpMV158, respectively. Nicking assays using supercoiled DNAs from pUB110, pDL287, and pE194 showed that MobM was able to relax, to a different degree, all plasmid DNAs. Our results suggest that cross-recognition of heterologous oriTs by Mob proteins could play an important role in the plasmid spreading between bacteria.Suggested Reviewers:Opposed Reviewers:Here we present information on the features of the MobM relaxase encoded by the promiscuous plasmid pMV158. The protein binds to oligonucleotides and to supercoiled plasmid DNA containing the origin of transfer. MobM-relaxed pMV158 DNA forms could be visualized by electron microscopy. The MobM protein relaxed supercoiled DNAs from other rolling circle-replicating plasmids which harbor origins of transfer with similarities to the one present in pMV158. Abstract 27The MobM relaxase (494 amino acids) encoded by the promiscuous 28 streptococcal plasmid pMV158 recognizes the plasmid origin of transfer, oriT pMV158 , 29 and converts supercoiled pMV158 DNA into relaxed molecules by cleavage of the 30 phosphodiester bond of a specific dinucleotide within the sequence 5"-GTGTG/TT-3" 31 ("/" being the nick site). After cleavage, the protein remains stably bound to the 5´-32 end of the nick site. Band-shift assays with single-stranded oligonucleotides and size-33 exclusion chromatography allowed us to show that MobM was able to generate 34 specific complexes with one of the inverted repeats of the oriT pMV158 , presumably 35 extruded as stem-loop structure. A number of tests have been performed to attain a 36 better characterization of the nicking activity of MobM and its linkage with its target 37 DNA. The optimal pH for DNA relaxa...

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Section: Recognition Of Heterologous Orits By Mobm Protein 420mentioning

confidence: 99%

Nicking activity of the pMV158 MobM relaxase on cognate and heterologous origins of transfer

Fernández-López

Lorenzo-Díaz

Pérez-Luque

et al. 2013

Plasmid

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“…The GϩC content in pTXL1 is 34%, in accordance with the GϩC percentage of Leuconostoc subsp. chromosomes and plasmids previously described (10,12). Surprisingly, sequence analysis revealed no similarities with genes encoding known proteins involved in replication, such as the replication initiation protein required for RCR plasmids.…”

mentioning

confidence: 99%

“…are a diverse group of heterofermentative LAB of considerable industrial importance. Many Leuconostoc species harbor one or more native plasmids of various sizes, but to date, only a few reports have dealt with RCR plasmids in species of the Leuconostoc genus (5,7,10,11,30) and none concerns identification of a non-RCR plasmid. The aim of the present study was to analyze the mode of replication of the Leuconostoc plasmid pTXL1 (6) in order to develop families of vectors designed for specific industrial applications.…”

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confidence: 99%

Identification of a Replicon from pTXL1, a Small Cryptic Plasmid from Leuconostoc mesenteroides subsp. mesenteroides Y110, and Development of a Food-Grade Vector

Biet

Cenatiempo

Fremaux³

2002

Appl Environ Microbiol

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A 2,665-bp cryptic plasmid, pTXL1, isolated from Leuconostoc mesenteroides subsp. mesenteroides Y110 was identified. This plasmid harbors a replicon localized on a 1,300-bp fragment. Two observations suggested that pTXL1 does not belong to rolling-circle replication (RCR)-type plasmids and most likely replicates via a theta mechanism. These hypotheses are supported by the observation that no detectable single-stranded intermediate was found for the replicon and that, unlike in RCR-type plasmids, the pTXL1 replicon sequence lacks an open reading frame encoding a replicase. The small-sized pTXL1 plasmid is stable and, according to its origin, can be considered in the "generally recognized as safe" category. Its ability to replicate in several lactic acid bacteria was exploited to develop a vector producing mesentericin Y105, a class II anti-Listeria bacteriocin. With this new vector, a recombinant industrial Leuconostoc cremoris strain able to produce mesentericin Y105 was constructed.

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“…The Cop protein is not found in pHPK255. Recently, a small cryptic plasmid pCI411 has been isolated from Leuconostoc lactis (Coffey et al, 1994). The sequence analysis of pCI411 reveals that this plasmid belongs to the pE194 family.…”

Section: Discussionmentioning

confidence: 99%

Bacillus stearothermophilus plasmid pSTK1 replicon is functional in Escherichia coli

et al. 1995

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A kanamycin-resistant plasmid possessing a thermostable replicon derived from Bacillus stearothermophilus cryptic plasmid pSTK1 was constructed. The plasmid could transform not only B. stearothermophilus and Bacillus subtilis, but also Gram-negative Escherichia coil The behavior of the plasmid in the hosts was examined. The plasmid was stably maintained even at 67°C in B. stearothermophilus without selective pressure. During the plasmid replication, single-stranded DNA (ssDNA) intermediates were found in E. coli, while these were not found in B. subtilis.

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Nucleotide sequence and structural organization of the small, broad-host-range plasmid pCI411 from Leuconostoc lactis 533

Cited by 42 publications

References 23 publications

Nicking activity of the pMV158 MobM relaxase on cognate and heterologous origins of transfer

Nicking activity of the pMV158 MobM relaxase on cognate and heterologous origins of transfer

Identification of a Replicon from pTXL1, a Small Cryptic Plasmid from Leuconostoc mesenteroides subsp. mesenteroides Y110, and Development of a Food-Grade Vector

Bacillus stearothermophilus plasmid pSTK1 replicon is functional in Escherichia coli

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