A small cryptic plasmid from Ruminobacter amylophilus NIAH-3 possesses functional mobilization properties

Ogata, K

doi:10.1016/s0378-1097(99)00512-1

Cited by 3 publications

(3 citation statements)

References 17 publications

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“…As previously shown, the pBBR1 oriT sequence looks like the RSA of several plasmids and mobilizable transposons from gram-positive bacteria (2,10,33). We have also found sequence similarities with other plasmids and transposons from gram-positive and gram-negative bacteria, and surprisingly, with a gene of the pUH1 plasmid described as a ␥-glutamyltranspeptidase gene (19) (a total of 35 sequences were found).…”

Section: Discussionsupporting

confidence: 71%

Mobilization Function of the pBHR1 Plasmid, a Derivative of the Broad-Host-Range Plasmid pBBR1

2001

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The pBHR1 plasmid is a derivative of the small (2.6-kb), mobilizable broad-host-range plasmid pBBR1, which was isolated from the gram-negative bacterium Bordetella bronchiseptica (R. Antoine and C. Locht, Mol. Microbiol. 6:1785-1799, 1992). Plasmid pBBR1 consists of two functional cassettes and presents sequence similarities with the transfer origins of several plasmids and mobilizable transposons from gram-positive bacteria. We show that the Mob protein specifically recognizes a 52-bp sequence which contains, in addition to the transfer origin, the promoter of the mob gene. We demonstrate that this gene is autoregulated. The binding of the Mob protein to the 52-bp sequence could thus allow the formation of a protein-DNA complex with a double function: relaxosome formation and mob gene regulation. We show that the Mob protein is a relaxase, and we located the nic site position in vitro. After sequence alignment, the position of the nic site of pBBR1 corresponds with those of the nick sites of the Bacteroides mobilizable transposon Tn4555 and the streptococcal plasmid pMV158. The oriT of the latter is characteristic of a family of mobilizable plasmids that are found in gram-positive bacteria and that replicate by the rolling-circle mechanism. Plasmid pBBR1 thus appears to be a new member of this group, even though it resides in gram-negative bacteria and does not replicate via a rolling-circle mechanism. In addition, we identified two amino acids of the Mob protein necessary for its activity, and we discuss their involvement in the mobilization mechanism.

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Section: Discussionsupporting

confidence: 71%

Mobilization Function of the pBHR1 Plasmid, a Derivative of the Broad-Host-Range Plasmid pBBR1

2001

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“…Recently, a novel class of rolling circle plasmids, the pHW126-like plasmids, was described (Rozhon et al, 2010). Currently, just four members of this group are known: pHW126, pIGRK and pIGMS31 (Smorawinska et al, 2012), which were isolated from Enterobacteriaceae, and the distantly related pRAO1 (Ogata et al, 1999), which was found in Ruminobacter amylophilus. These plasmids are characterized by low G + C contents of 32-40% and small sizes of < 3 kb.…”

Section: Introductionmentioning

confidence: 99%

Identification of the region required for maintaining pHW126 in its monomeric form

Rozhon

Khan

Poppenberger

2012

FEMS Microbiol Lett

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The pHW126‐like plasmids are a recently discovered small group of cryptic plasmids replicating by the rolling circle mode. The replication origin of pHW126 consists of a conserved stretch, four perfect direct repeats and a so‐called accessory region. The latter increases plasmid stability but is not absolutely necessary for replication. Here, we report that deletion of the accessory region causes rapid multimerization of pHW126. While the number of pHW126‐units per cell remains constant, the number of physically independent plasmid molecules is reduced by approximately 40%, rendering random distribution to daughter cells less effective. A conserved inverted repeat within the accessory region could be identified as a sequence necessary for maintaining pHW126 in its monomeric form. A mutant version of pHW126 lacking this inverted repeat could be rescued by placing the single‐strand initiation site (ssi) of pHW15 on the plus strand, while including the ssi in the opposite direction had no effect. Thus, our data provide evidence that multimer formation is, besides copy number reduction and ssDNA accumulation, an additional means how loss of a mechanism ensuring efficient lagging strand synthesis may cause destabilization of rolling circle plasmids.

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“…These modules also occur in some plasmids from Gram-negative hosts, e.g. those from Bordetella bronchiseptica (Antoine & Locht, 1992) and Ruminobacter amylophilus (Ogata et al, 1999). To confirm the presence of such modules in pWKS1, a detailed functional analysis of the plasmid was carried out in a later part of this study.…”

Section: Nucleotide Sequence Of Pwks1mentioning

confidence: 99%

Molecular characterization of functional modules of plasmid pWKS1 of Paracoccus pantotrophus DSM 11072 The GenBank accession number for the sequence reported in this paper is AF482428.

et al. 2002

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The complete nucleotide sequence of the small, cryptic plasmid pWKS1 (2697 bp) of Paracoccus pantotrophus DSM 11072 was determined. The GMC content of the sequence of this plasmid was 62 mol %. Analysis revealed that over 80 % of the plasmid genome was covered by two ORFs, ORF1 and ORF2, which were capable of encoding putative peptides of 441 and 378 kDa, respectively. Mutational analysis showed that ORF2 was crucial for plasmid replication.

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A small cryptic plasmid from Ruminobacter amylophilus NIAH-3 possesses functional mobilization properties

Cited by 3 publications

References 17 publications

Mobilization Function of the pBHR1 Plasmid, a Derivative of the Broad-Host-Range Plasmid pBBR1

Mobilization Function of the pBHR1 Plasmid, a Derivative of the Broad-Host-Range Plasmid pBBR1

Identification of the region required for maintaining pHW126 in its monomeric form

Molecular characterization of functional modules of plasmid pWKS1 of Paracoccus pantotrophus DSM 11072 The GenBank accession number for the sequence reported in this paper is AF482428.

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