A DNA sequence outside the pUB110 minimal replicon is required for normal replication in<i>Bacillus subtilis</i>

Viret, Jean-François; Alonso, Juan C.

doi:10.1093/nar/16.10.4389

Cited by 40 publications

(54 citation statements)

References 26 publications

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“…The use of indirect assays which correlate M-O activity with transformability into a dnaD23 mutant B. subtilis strain (39) rather than a direct measure of ssDNA of deletion derivatives as presented here may explain the discrepancy. Concomitant deletion of another locus on the plasmid, e.g., the pre locus which maps adjacent to M-O (12), may have affected the previously published results (39).…”

Section: Discussionmentioning

confidence: 92%

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Replication origins of single-stranded-DNA plasmid pUB110

et al. 1989

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The two replication origins of plasmid pUB110 have been characterized. The site of initiation of DNA replication at the plus origin was mapped to within an 8-base-pair sequence. DNA synthesis initiated at the origin was made to terminate precociously in an inserted sequence of 18 base pairs that is homologous to a sequence in the origin. This suggests that pUB110 replicates as a rolling circle. The minus origin of plasmid pUB10 has been characterized, and the minimal sequence required for function has been determined. As with other minus origins, activity is orientation specific with respect to the direction of replication. Its activity is sensitive to rifampin in vivo, suggesting that RNA polymnerase catalyzes single-strand to double-strand conversion. Unlike all other plasmids of gram-positive bacteria thus far described, the pUB110 minus origin is functional in more than one host.

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Section: Discussionmentioning

confidence: 92%

“…Concomitant deletion of another locus on the plasmid, e.g., the pre locus which maps adjacent to M-O (12), may have affected the previously published results (39).…”

Section: Discussionmentioning

confidence: 99%

Replication origins of single-stranded-DNA plasmid pUB110

et al. 1989

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“…Surprisingly, dnaD23 cells harbouring the pDG148 vector alone presented an exacerbated temperature sensitivity, as they could not grow at 42∞C (dnaD23 cells can sustain growth up to 45∞C). We interpret this defect as a titration of DnaD by the plasmid, because its rolling-circle replication is known to depend on DnaD (Viret and Alonso, 1988). In contrast, when DnaD23 was overexpressed in dnaD23 cells from pSMG39, cells were viable at 42∞C.…”

Section: Dnad23 Is a 'Loss-of-function' Mutationmentioning

confidence: 99%

Functional interplay between the Bacillus subtilis DnaD and DnaB proteins essential for initiation and re‐initiation of DNA replication

Bruand

Velten

McGovern

et al. 2005

Molecular Microbiology

102

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SummaryInitiation and re-initiation of chromosomal DNA replication in bacteria rely on divergent multiprotein assemblies, which direct the functional delivery of the replicative helicase on single-stranded DNA (ssDNA) at specific sites. These two processes are triggered either at the single chromosomal origin oriC or at arrested forks by the conserved DnaA and PriA proteins respectively. In Bacillus subtilis , these two pathways further require the three essential proteins DnaB, DnaD and DnaI, restrictively encoded in Gram positive bacteria of low GC content. We have recently shown that DnaI and DnaB act as a pair of loaders of the DnaC replicative helicase. The role of DnaD appeared more enigmatic. It was previously shown to interact with DnaA and to display weak ssDNA binding activity. Here, we report that purified DnaD can interact physically with PriA and with DnaB. We show that the lethality of the temperature-sensitive dnaD23 mutant can be suppressed by different DnaB point mutants, which were found to be identical to the suppressors of priA null mutants. The DnaD23 protein displays lower ssDNA binding activity than DnaD. Conversely, the DnaB75 protein, the main dnaD23 suppressor, has gained affinity for ssDNA. Finally, we observed that this interplay between DnaD and DnaB is crucial for their concerted interaction with SSBcoated ssDNA, which is the expected substrate for the loading of the replicative helicase in vivo . Altogether, these results highlight the need for both DnaD and DnaB to interact individually and together with ssDNA during the early stages of initiation and re-initiation of chromosomal DNA replication. They also point at a main structural role of DnaD in the multiprotein assemblies built during these two essential processes.

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“…The ssoU is the most promiscuous sso origin characterized so far, as it seems to be fully functional in many, if not all, Firmicutes (121,(139)(140)(141)(142). The ssoU origin was first identified in staphylococcal plasmid pUB110 (142), within a ∼250-bp DNA fragment with the potential to form several hairpin structures containing symmetric and asymmetric internal loops in addition to the terminal one (139).…”

Section: The Single-strand Originmentioning

confidence: 99%

“…The ssoU origin was first identified in staphylococcal plasmid pUB110 (142), within a ∼250-bp DNA fragment with the potential to form several hairpin structures containing symmetric and asymmetric internal loops in addition to the terminal one (139). Sequences nearly identical to the pUB110-ssoU were subsequently found in the streptococcal pMV158 and Bacillus pTB913 plasmids, and the involvement of these elements in lagging-strand synthesis was also proved (143).…”

Section: The Single-strand Originmentioning

confidence: 99%

Plasmid Rolling-Circle Replication

et al. 2015

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Plasmids are DNA entities that undergo controlled replication independent of the chromosomal DNA, a crucial step that guarantees the prevalence of the plasmid in its host. DNA replication has to cope with the incapacity of the DNA polymerases to start de novo DNA synthesis, and different replication mechanisms offer diverse solutions to this problem. Rolling-circle replication (RCR) is a mechanism adopted by certain plasmids, among other genetic elements, that represents one of the simplest initiation strategies, that is, the nicking by a replication initiator protein on one parental strand to generate the primer for leading-strand initiation and a single priming site for lagging-strand synthesis. All RCR plasmid genomes consist of a number of basic elements: leading strand initiation and control, lagging strand origin, phenotypic determinants, and mobilization, generally in that order of frequency. RCR has been mainly characterized in Gram-positive bacterial plasmids, although it has also been described in Gram-negative bacterial or archaeal plasmids. Here we aim to provide an overview of the RCR plasmids' lifestyle, with emphasis on their characteristic traits, promiscuity, stability, utility as vectors, etc. While RCR is one of the best-characterized plasmid replication mechanisms, there are still many questions left unanswered, which will be pointed out along the way in this review.

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A DNA sequence outside the pUB110 minimal replicon is required for normal replication inBacillus subtilis

Cited by 40 publications

References 26 publications

Replication origins of single-stranded-DNA plasmid pUB110

Replication origins of single-stranded-DNA plasmid pUB110

Functional interplay between the Bacillus subtilis DnaD and DnaB proteins essential for initiation and re‐initiation of DNA replication

Plasmid Rolling-Circle Replication

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