The Promiscuous Plasmids pIP501 and pAMβ1 from Gram-Positive Bacteria Encode Complementary Resolution Functions

Pujol, Catherine; Ehrlich, S. Dusko; Janniere, Laurent

doi:10.1006/plas.1994.1010

Cited by 27 publications

(26 citation statements)

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“…To compare better the three plasmids, the region located downstream of resIP was sequenced. The 5552 bp sequence reported here (accession number U00453) forms, with the previously published sequences, a contig of 9905 bp (Brantl et al, 1990;Trieu-Cuot et al, 1992;Pujol et al, 1994). The 5552 bp are strongly (> 90%) homologous to the corresponding regions of pAM␤1 and pSM19035, with the notable exception of the central part where an insert of 2893 bp was found.…”

Section: Detection Of a Rc Plasmid In Pip501supporting

confidence: 58%

“…pIP501, a member of the pAM␤1/pSM19035 family, has been shown previously to carry a resolution system (res/ resIP ) highly homologous to that of pAM␤1 downstream of repR (Pujol et al, 1994). Further downstream, and in different contigs, genes encoding chloramphenicol (Cm) resistance (cat ), inducible MLS resistance (erm1-erm2 respectively) and a ␦-like protein have been described (Evans and Macrina, 1983;Trieu-Cuot et al, 1992;Langella et al, 1993;Brantl et al, 1994).…”

Section: Detection Of a Rc Plasmid In Pip501mentioning

confidence: 99%

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Inhibition of a naturally occurring rolling‐circle replicon in derivatives of the theta‐replicating plasmid pIP501

Pujol

Chédin

Ehrlich

et al. 1998

Molecular Microbiology

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SummaryThe mechanisms ensuring regulation of DNA replication in genomes containing multiple replicons are poorly understood. In this report, we addressed this question by analysing in Bacillus subtilis the replication of a derivative of the promiscuous plasmid pIP501 that carries a rolling-circle and a theta replicon. Genetic analyses revealed that the rolling-circle replicon is strongly inhibited in the derivative and that inhibition requires three elements involved in theta replication: the replication origin, the initiator RepR protein and strong transcription of the repR gene. Inhibition is, however, independent of DNA synthesis at the theta origin. We conclude that rolling-circle inhibition is caused by an inhibitor y signal encoded by the theta replicon and propose that the signal is composed, at least, of the RepR protein bound to its cognate origin.

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Section: Detection Of a Rc Plasmid In Pip501supporting

confidence: 58%

Section: Detection Of a Rc Plasmid In Pip501mentioning

confidence: 99%

Inhibition of a naturally occurring rolling‐circle replicon in derivatives of the theta‐replicating plasmid pIP501

Pujol

Chédin

Ehrlich

et al. 1998

Molecular Microbiology

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“…In pAM␤1-related plasmids, this sequence is a res site which binds a resolvase almost identical to Res␤ (Pujol et al, 1994;Rojo and Alonso, 1994). In plasmids of the ColE1 family, the sequence is an origin of transfer, bom, known to be the target for several proteins required for mobilization (Mob; Veltkamp and Stuitje, 1981;Morlon et al, 1988;Boyd et al, 1989;Fu et al, 1995).…”

Section: Discussionmentioning

confidence: 99%

“…Such a switch may take place in plasmids of the pAM␤1 and ColE1 families because they all carry a primosome-assembly site on the lagging-strand template, immediately upstream of a protein-binding site (Nomura and Ray, 1980;Marians et al, 1982;Nijkamp et al, 1986;Morlon et al, 1988;Boyd et al, 1989;Brantl et al, 1990;Pujol et al, 1994;Bruand et al, 1995;Fu et al, 1995). The arrested D-loop intermediates that may form in these plasmids through the arrest of Pol I at a Res-res or Mob-bom roadblock could therefore be the substrate for Pol I to Pol III HE switching and for replisome entry.…”

Section: Discussionmentioning

confidence: 99%

“…Plasmid pIL253 was described elsewhere (Bruand et al, 1993). pHV1461 is a kanamycin-resistant pUB110 derivative which produces Res␤ under the control of the IPTG-inducible spac-I promoter (Pujol et al, 1994). The pHP727FX plasmid mainly contains the pBR322 and f1(þ) replication origins, a 310-bp-long inverted repeat derived from Tn5 ends and an erythromycin determinant (Canceill and Ehrlich, 1996).…”

Section: Bacterial Strains and Plasmidsmentioning

confidence: 99%

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Replication terminus for DNA polymerase I during initiation of pAMβ1 replication: role of the plasmid‐encoded resolution system

Janniere

Bidnenko

McGovern

et al. 1997

Molecular Microbiology

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SummaryReplication of plasmid pAM␤1 is initiated by DNA polymerase I (Pol I) and completed by DNA polymerase III holoenzyme contained in the replisome machinery. In this study we report that initiation of DNA replication generates D-loop structures containing the nascent leading strand paired to its template, and that D-loop extension is arrested Ϸ230 bp from the initiation site of DNA synthesis in the presence of the plasmid-encoded resolvase. In vitro and in vivo data suggest that this arrest is caused by a collision between Pol I and the resolvase bound to its target. As the arrested D-loop replication intermediates carry a single-stranded primosome-assembly site, we hypothesize that the biological role of the replication arrest is to limit the region replicated by Pol I and to promote the replacement of Pol I by the replisome in order to initiate concerted synthesis of the leading and lagging strands.

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Primosome assembly site in Bacillus subtilis.

Bruand¹,

Ehrlich²,

Janniere³

1995

The EMBO Journal

104

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Corresponding author A single-strand initiation site was detected on the Enterococcus faecalis plasmid pAMP1 by its ability to prevent accumulation of single stranded DNA of a rolling circle plasmid, both in Bacillus subtilis and Staphylococcus aureus. This site, designated ssiA, is located on the lagging strand template, -150 bp downstream from the replication origin. ssiA priming activity requires the DnaE primase, the DnaC replication fork helicase, as well as the products of the dnaB, dnaD and dnal genes of B.subtilis, but not the RNA polymerase. The primase and the replication fork helicase requirements indicate that ssiA is a primosome assembly site.Interestingly, the pAM01 lagging strand synthesis is inefficient when any of the proteins involved in ssiA activity is mutated, but occurs efficiently in the absence of ssiA. This suggests that normal plasmid replication requires primosome assembly and that the primosome can assemble not only at ssiA but also elsewhere on the plasmid. This work for the first time describes a primosome in a Gram-positive bacterium. Involvement of the B.subtilis proteins DnaB, DnaD and DnaI, which do not have any known analogue in Escherichia coli, raises the possibility that primosome assembly and/or function in B.subtilis differs from that in E.coli.

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The Promiscuous Plasmids pIP501 and pAMβ1 from Gram-Positive Bacteria Encode Complementary Resolution Functions

Cited by 27 publications

References 0 publications

Inhibition of a naturally occurring rolling‐circle replicon in derivatives of the theta‐replicating plasmid pIP501

Inhibition of a naturally occurring rolling‐circle replicon in derivatives of the theta‐replicating plasmid pIP501

Replication terminus for DNA polymerase I during initiation of pAMβ1 replication: role of the plasmid‐encoded resolution system

Primosome assembly site in Bacillus subtilis.

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