Definition and polarity of action of DNA replication terminators in Bacillus subtilis

Smith, Marc D.; Wake, R.G.

doi:10.1016/0022-2836(92)90214-5

Cited by 52 publications

(81 citation statements)

References 23 publications

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“…B. subtilis SU230, a constitutive overproducer of RTP, was used for in vivo replication fork arrest assays (16).…”

Section: Methodsmentioning

confidence: 99%

“…Plasmids-pID1 was constructed by ligating the annealed oligonucleotides, 5Ј-AATTCTATGAACATAATGTTCATAG and 5Ј-GATCCTA-TGAACATTATGTTCATAG, to the large EcoRI-BamHI fragment of pWS64-1 (16). pID6 was constructed by ligating the annealed oligonucleotides, 5Ј-AATTCGGCTGCAGAACTAAGAAAACTATGAACATAAT-GTTCATAG and 5Ј-GATCCTATGAACATTATGTTCATAGTTTTCTTA-GTTCTGCAGCCG, to the large EcoRI-BamHI fragment of pWS64-1.…”

Section: Methodsmentioning

confidence: 99%

“…Two synthetic double-stranded oligonucleotides (one for the "distal" and one for the "proximal" half-site, listed respectively after the pTer moiety of the plasmid name; see Fig. 3) were ligated in vitro to form the full-length sequence, and the product was ligated to the large BamHI-SalI fragment of pWS64-1 (16). After transformation and purification from E. coli, all plasmids were sequence-verified over the region of the cloned terminator sequences.…”

Section: Methodsmentioning

confidence: 99%

“…The polar Ter sites contain low and high affinity half-sites (A and B sites, respectively) that each bind one dimer, and the level of protein-DNA contact is significantly higher at the B site (13). A replication fork is arrested only when it approaches the B site side of the complex, although a single RTP dimer bound to an isolated B site cannot cause arrest (16). It is clear that interaction between the two bound RTP dimers (13,17,18), possibly involving direct protein-protein contacts (14), is essential for fork arrest at Ter sites.…”

mentioning

confidence: 99%

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A Complex Mechanism Determines Polarity of DNA Replication Fork Arrest by the Replication Terminator Complex of Bacillus subtilis

Duggin

Matthews

Dixon

et al. 2005

Journal of Biological Chemistry

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Two dimers of the replication terminator protein (RTP) of Bacillus subtilis bind to a chromosomal DNA terminator site to effect polar replication fork arrest. Cooperative binding of the dimers to overlapping halfsites within the terminator is essential for arrest. It was suggested previously that polarity of fork arrest is the result of the RTP dimer at the blocking (proximal) side within the complex binding very tightly and the permissive-side RTP dimer binding relatively weakly. In order to investigate this "differential binding affinity" model, we have constructed a series of mutant terminators that contain half-sites of widely different RTP binding affinities in various combinations. Although there appeared to be a correlation between binding affinity at the proximal half-site and fork arrest efficiency in vivo for some terminators, several deviated significantly from this correlation. Some terminators exhibited greatly reduced binding cooperativity (and therefore have reduced affinity at each half-site) but were highly efficient in fork arrest, whereas one terminator had normal affinity over the proximal half-site, yet had low fork arrest efficiency. The results show clearly that there is no direct correlation between the RTP binding affinity (either within the full complex or at the proximal half-site within the full complex) and the efficiency of replication fork arrest in vivo. Thus, the differential binding affinity over the proximal and distal half-sites cannot be solely responsible for functional polarity of fork arrest. Furthermore, efficient fork arrest relies on features in addition to the tight binding of RTP to terminator DNA.Replication terminator proteins are a diverse group of DNAbinding proteins that cause DNA replication fork arrest (or pausing) at designated DNA terminator sites. They have roles in a variety of tasks, including the coordination of DNA replication and transcription at eukaryotic rDNA loci (1, 2), regulation of mating-type switching in yeast by strand-specific imprinting during replication (3, 4), and participation in the final stages of bacterial chromosome replication and partitioning (5).These functions require the DNA replication fork to be blocked at the terminator site from only one direction of approach. Thus, a terminator protein-DNA complex exhibits functional polarity. An understanding of the mechanism of polar replication fork arrest has remained elusive.Considerable progress in the characterization of replication termination systems has been made with those from Bacillus subtilis and Escherichia coli (6, 7), and this has recently aided the study of analogous eukaryotic systems (4, 8, 9). In both bacteria, the numerous terminator (Ter) 1 sites are clustered in the terminal region of the chromosome and are arranged so as to control the location of replication fork fusion at the end of a round of replication (for a review see Ref. 10). Despite the same apparent function of these two bacterial systems, the terminator proteins from E. coli and B. subtilis share no sequ...

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“…B. subtilis SU230, a constitutive overproducer of RTP, was used for in vivo replication fork arrest assays (16).…”

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

See 2 more Smart Citations

A Complex Mechanism Determines Polarity of DNA Replication Fork Arrest by the Replication Terminator Complex of Bacillus subtilis

Duggin

Matthews

Dixon

et al. 2005

Journal of Biological Chemistry

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“…In B. subtilis, as well as E. coli, the termination complex blocks DNA replication in a polar manner. Replication forks that encounter the B. subtilis RTP-Ter complex from the core side are blocked, although those that encounter RTP-Ter from the auxiliary side are not (8). The termination complexes of B. subtilis and E. coli inhibit helicase-mediated DNA unwinding (9,10), although the mechanism of helicase inhibition is still debated (11).…”

mentioning

confidence: 99%

Effects of replication termination mutants on chromosome partitioning in Bacillussubtilis

Lemon

Kurtser²,

Grossman³

2000

Proceedings of the National Academy of Sciences

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Many circular genomes have replication termination systems, yet disruption of these systems does not cause an obvious defect in growth or viability. We have found that the replication termination system of Bacillus subtilis contributes to accurate chromosome partitioning. Partitioning of the terminus region requires that chromosome dimers, that have formed as a result of RecA-mediated homologous recombination, be resolved to monomers by the site-specific recombinase encoded by ripX. In addition, the chromosome must be cleared from the region of formation of the division septum. This process is facilitated by the spoIIIE gene product which is required for movement of a chromosome out of the way of the division septum during sporulation. We found that deletion of rtp, which encodes the replication termination protein, in combination with mutations in ripX or spoIIIE, led to an increase in production of anucleate cells. This increase in production of anucleate cells depended on recA, indicating that there is probably an increase in chromosome dimer formation in the absence of the replication termination system. Our results also indicate that SpoIIIE probably enhances the function of the RipX recombinase system. We also determined the subcellular location of the replication termination protein and found that it is a good marker for the position of the chromosome terminus.

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Termination ofDNAReplication in Prokaryotes

Rudolph

Corocher

Grainge

et al. 2019

Encyclopedia of Life Sciences

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Most bacteria and archaea have circular chromosomes, in which DNA replication begins at a site known as an origin of replication. Double-stranded DNA unwound at the origin creates two replication forks that are engaged by DNA polymerase complexes (replisomes) that advance each fork and proceed in 21 opposite directions away from the origin, copying the original strands. Termination of DNA replication 22 occurs when the two forks meet and fuse, creating two separate double-stranded DNA molecules. In 23 the well-studied bacteria Escherichia coli and Bacillus subtilis, this occurs in the terminus region, which is situated diametrically opposite the origin. Failure to terminate chromosome replication correctly can lead to problems with genome function and stability, including DNA over-replication. In contrast, some 26 archaea have multi-origin chromosomes and do not appear to specifically regulate the location of termination.

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Definition and polarity of action of DNA replication terminators in Bacillus subtilis

Cited by 52 publications

References 23 publications

A Complex Mechanism Determines Polarity of DNA Replication Fork Arrest by the Replication Terminator Complex of Bacillus subtilis

A Complex Mechanism Determines Polarity of DNA Replication Fork Arrest by the Replication Terminator Complex of Bacillus subtilis

Effects of replication termination mutants on chromosome partitioning in Bacillussubtilis

Termination ofDNAReplication in Prokaryotes

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