All Major Regions of FtsK Are Required for Resolution of Chromosome Dimers

Boyle, David S.; Grant, D.A.D.; Draper, G. Craig; Donachie, W.

doi:10.1128/jb.182.14.4124-4127.2000

Cited by 15 publications

(12 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…least one other protein, FtsK, is required for resolution of chromosome multimers in vivo (10,37). B. subtilis SpoIIIE and YtpT are, respectively, 48 and 49% identical to the C terminus of the E. coli FtsK protein.…”

Section: Deletion Induced Filamentationmentioning

confidence: 99%

Identification and Characterization of the dif Site from Bacillus subtilis

2001

View full text Add to dashboard Cite

Bacteria with circular chromosomes have evolved systems that ensure multimeric chromosomes, formed by homologous recombination between sister chromosomes during DNA replication, are resolved to monomers prior to cell division. The chromosome dimer resolution process in Escherichia coli is mediated by two tyrosine family site-specific recombinases, XerC and XerD, and requires septal localization of the division protein FtsK. The Xer recombinases act near the terminus of chromosome replication at a site known as dif (Ecdif). In Bacillus subtilis the RipX and CodV site-specific recombinases have been implicated in an analogous reaction. We present here genetic and biochemical evidence that a 28-bp sequence of DNA (Bsdif), lying 6°counter-clockwise from the B. subtilis terminus of replication (172°), is the site at which RipX and CodV catalyze site-specific recombination reactions required for normal chromosome partitioning. Bsdif in vivo recombination did not require the B. subtilis FtsK homologues, SpoIIIE and YtpT. We also show that the presence or absence of the B. subtilis SP␤-bacteriophage, and in particular its yopP gene product, appears to strongly modulate the extent of the partitioning defects seen in codV strains and, to a lesser extent, those seen in ripX and dif strains.Cells with circular chromosomes and homologous recombination systems must be able to resolve chromosome dimers, or higher-order multimeric forms, that are generated by an odd number of recombination events between sister chromosomes during DNA replication. Failure to resolve chromosome dimers into monomers will prevent the proper partitioning of genomic material to newly forming daughter cells. A model for the coordination of chromosome dimer resolution and cell division has been elaborated in Escherichia coli based on a substantial accumulation of in vivo and in vitro data. In E. coli, two tyrosine family site-specific recombinases, XerC and XerD, act in concert at a site near the terminus of chromosome replication known as dif to resolve chromosome dimers into monomers during the process of cell division (9,6,12,22). Deletion of Ecdif or mutations in xerC or xerD result in the development of a subpopulation of filamentous cells containing abnormally partitioned nucleoids. In addition, it has been demonstrated that the FtsK protein must be located at the constricting septum for the Xer-mediated resolution of chromosomes to occur (26,37,39). Similarly, plasmids containing the 28-bp minimal Ecdif site show dependence on FtsK for Xer-mediated inter-and intramolecular site-specific recombination in vivo (33). Dimeric chromosomes are thought to arise primarily as a by-product of homologous recombination events that enable bacteria to re-initiate replication at stalled replication forks. Although sister chromatid exchanges that generate dimers have been estimated to occur in approximately 15% of cells within a growing population, it is currently thought that recombinational DNA repair of stalled replication forks is a major housekeeping event...

show abstract

Section: Deletion Induced Filamentationmentioning

confidence: 99%

Identification and Characterization of the dif Site from Bacillus subtilis

2001

View full text Add to dashboard Cite

show abstract

“…Finally, the putative nucleotide binding site of FtsK was critical for its role in Xer recombination. It had been previously reported that two FtsK N-terminal truncations (starting at residue 825 and 863) fused to a polyhistidine tag (Boyle et al 2000), and a protein containing domain 2 and 3 of FtsK did not complement Xer recombination at dif. However, we believe that these negative results may reflect a problem of expression or a lack of activity of the tagged proteins.…”

Section: The C-terminal Domain Of Ftsk Is Sufficient To Support a Commentioning

confidence: 99%

FtsK functions in the processing of a Holliday junction intermediate during bacterial chromosome segregation

Barre¹,

Aroyo²,

Colloms³

et al. 2000

Genes Dev.

104

145

View full text Add to dashboard Cite

In bacteria with circular chromosomes, homologous recombination can generate chromosome dimers that cannot be segregated to daughter cells at cell division. Xer site-specific recombination at dif, a 28-bp site located in the replication terminus region of the chromosome, converts dimers to monomers through the sequential action of the XerC and XerD recombinases. Chromosome dimer resolution requires that dif is positioned correctly in the chromosome, and the activity of FtsK, a septum-located protein that coordinates cell division with chromosome segregation. Here, we show that cycles of XerC-mediated strand exchanges form and resolve Holliday junction intermediates back to substrate irrespective of whether conditions support a complete recombination reaction. The C-terminal domain of FtsK is sufficient to activate the exchange of the second pair of strands by XerD, allowing both intra-and intermolecular recombination reactions to go to completion. Proper positioning of dif in the chromosome and of FtsK at the septum is required to sense the multimeric state of newly replicated chromosomes and restrict complete Xer reactions to dimeric chromosomes.

show abstract

“…The 11-bp insertion is located in the second open reading frame (ORF) and contains an in-frame stop codon. The fourth ORF of this cluster encodes a protein with FtsK motifs that may be involved in cell division (2,3,4). The fifth ORF of this operon has already been identified as a M. avium subsp.…”

mentioning

confidence: 99%

Characterization of Genetic Differences between Mycobacterium avium subsp. paratuberculosis Type I and Type II Isolates

et al. 2003

View full text Add to dashboard Cite

A combination of representational difference analysis and comparative DNA sequencing revealed that four type I (sheep) isolates of Mycobacterium avium subsp. paratuberculosis were differentiated from nine type II (bovine) isolates by the presence of an 11-bp insertion in a novel M. avium subsp. paratuberculosis-specific region of genomic DNA. Further, our studies show that M. avium subsp. paratuberculosis type I isolates contain three type-specific loci that are missing in M. avium subsp. paratuberculosis type II but are present in M. avium subsp. avium. Taken together, the results are consistent with the hypothesis that M. avium subsp. paratuberculosis type I strains are an evolutionary intermediate between M. avium subsp. avium and M. avium subsp. paratuberculosis type II isolates or share a subset of M. avium subsp. avium type-specific loci through horizontal transfer.Mycobacterium avium subsp. paratuberculosis is the causative agent of paratuberculosis (Johne's diseases), a chronic granulomatous enteritis in ruminants. The disease is prevalent in domestic and wild animals worldwide (15,16,24,31) and has a considerable economic impact on the livestock industry (13). In humans, M. avium subsp. paratuberculosis has been suggested to be involved in Crohn's disease (6,11,20). M. avium subsp. paratuberculosis shows a DNA homology of more than 90% to M. avium subsp. avium (14,26). It differs from other subspecies of M. avium by its dependence on the iron chelator mycobactin for growth in culture (9, 30) and by the presence of multiple copies of the insertion sequences IS900 (12) and ISMav2 (29).By using analyses by pulsed-field electrophoresis (PFGE) and IS900-restriction fragment length polymorphisms, M. avium subsp. paratuberculosis isolates have been divided into two distinct types, with type I comprising very slow-growing, mostly pigmented isolates forming smooth and uniform colonies mainly obtained from sheep and other small ruminants and type II comprising slow-growing, nonpigmented isolates forming rough and nonuniform colonies exhibiting a very broad host range (27). Both types can also be differentiated by IS1311-PCR (19) and by a specific multiplex PCR (8). However, the molecular differences between M. avium subsp. paratuberculosis type I and type II strains which might give important clues with respect to (i) the evolutionary relationship between the two M. avium subsp. paratuberculosis types, (ii) the different phenotypes, and (iii) the differences in host preference have not as yet been elucidated.In this study we investigated the differences between M. avium subsp. paratuberculosis type I and type II strains by applying the technique of representational difference analysis (RDA) (18) to isolate M. avium subsp. paratuberculosis subspecies as well as type-specific DNA fragments. The isolation was followed by PCR, restriction endonuclease digest, and nucleotide sequence analyses of the respective fragments obtained from M. avium subsp. paratuberculosis isolated from various hosts.The bacterial strains an...

show abstract

All Major Regions of FtsK Are Required for Resolution of Chromosome Dimers

Cited by 15 publications

References 12 publications

Identification and Characterization of the dif Site from Bacillus subtilis

Identification and Characterization of the dif Site from Bacillus subtilis

FtsK functions in the processing of a Holliday junction intermediate during bacterial chromosome segregation

Characterization of Genetic Differences between Mycobacterium avium subsp. paratuberculosis Type I and Type II Isolates

Contact Info

Product

Resources

About