Behavior of Sister Copies of Mini-F Plasmid after Synchronized Plasmid Replication in<i>Escherichia coli</i>Cells

Onogi, Toshinari; Miki, Takeyoshi; Hiraga, Sota

doi:10.1128/jb.184.11.3142-3145.2002

Cited by 41 publications

(33 citation statements)

References 18 publications

(28 reference statements)

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“…When the production rates and the decay rates of SopA, SopB and the nucleation core were all reduced in the simulation, the number of SopB multimer peaks reduced but these peaks were positioned in an ordered manner. This is consistent with the results described by Onogi et al, 34 in which reduced copies of temperature-sensitive replication initiation mutant of mini-F plasmid localized at ordered positions. In the absence of SopA, SopB localized at mid-cell or pole-proximal region without formation of spiral structures (data not shown).…”

Section: Discussionsupporting

confidence: 93%

Subcellular Positioning of F Plasmid Mediated by Dynamic Localization of SopA and SopB

Adachi

Hori

Hiraga

2006

Journal of Molecular Biology

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Section: Discussionsupporting

confidence: 93%

Subcellular Positioning of F Plasmid Mediated by Dynamic Localization of SopA and SopB

Adachi

Hori

Hiraga

2006

Journal of Molecular Biology

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“…Establishment of replisome position. It is clear from several studies that the active replication machinery occupies characteristic subcellular locations in a variety of bacterial species (3,6,8,20,24,28,33,34,49,50,54,55). It is not clear how the position of the replisome is established or maintained.…”

Section: Resultsmentioning

confidence: 99%

“…During replication, chromosomal DNA moves to the replication factory, is duplicated, and then moves away from the central factory (34). Replication in E. coli also appears to take place at or near midcell or at positions that will be midcell (3,8,28,49,50,54,55). In contrast, in Caulobacter crescentus, the replication factory is initially positioned at the stalked cell pole and gradually moves to midcell as replication proceeds (24).…”

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confidence: 99%

“…Many plasmids encode partitioning systems that contribute to the stability and subcellular positioning of the plasmid. For example, in E. coli, both the unit copy plasmid F and plasmid prophage P1 are found predominantly at midcell or the cell quarters, and this positioning depends on a functional partitioning system named Par in P1 and Sop (stability of plasmids) in F (12,14,19,37,44,50).…”

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Multicopy Plasmids Affect Replisome Positioning inBacillus subtilis

et al. 2004

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The DNA replication machinery, various regions of the chromosome, and some plasmids occupy characteristic subcellular positions in bacterial cells. We visualized the location of a multicopy plasmid, pHP13, in living cells of Bacillus subtilis using an array of lac operators and LacI-green fluorescent protein (GFP). In the majority of cells, plasmids appeared to be highly mobile and randomly distributed. In a small fraction of cells, there appeared to be clusters of plasmids located predominantly at or near a cell pole. We also monitored the effects of the presence of multicopy plasmids on the position of DNA polymerase using a fusion of a subunit of DNA polymerase to GFP. Many of the plasmid-containing cells had extra foci of the replisome, and these were often found at uncharacteristic locations in the cell. Some of the replisome foci were dynamic and highly mobile, similar to what was observed for the plasmid. In contrast, replisome foci in plasmid-free cells were relatively stationary. Our results indicate that in B. subtilis, plasmid-associated replisomes are recruited to the subcellular position of the plasmid. Extending this notion to the chromosome, we postulated that the subcellular position of the chromosomally associated replisome is established by the subcellular location of oriC at the time of initiation of replication.The DNA replication machinery and regions of chromosomal DNA occupy characteristic positions within bacterial cells (reviewed in references 15, 17, 35, and 56). In Bacillus subtilis, the origin of chromosomal replication, oriC, of the single circular chromosome is typically positioned at or near midcell in cells with a single unreplicated origin and at or near the cell quarters (future midcell) in growing cells that contain a partly duplicated chromosome (10,31,32,36,40,57,58,61,62). In Escherichia coli, the location of sister oriC regions appears to be at the cell quarters or closer to the cell poles (14,29,38,45,47,54,55).Replication of the B. subtilis chromosome typically takes place in centralized replication factories. The replisome (the multiprotein complex that includes the replicative polymerase, helicase, and associated proteins that are present at the replication fork) is localized at or near midcell or at the cell quarters which will be midcell following division (33,34). During replication, chromosomal DNA moves to the replication factory, is duplicated, and then moves away from the central factory (34). Replication in E. coli also appears to take place at or near midcell or at positions that will be midcell (3,8,28,49,50,54,55). In contrast, in Caulobacter crescentus, the replication factory is initially positioned at the stalked cell pole and gradually moves to midcell as replication proceeds (24).The mechanisms that establish and maintain the subcellular position of the replisome are not known, nor is it known how the location of oriC is established or maintained, although several genes and sites are known to contribute to positioning of the origin region (4,13,15,17,25,...

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“…The finding that DNA replication occurs in stationary factories in the regions of the mid-cell and quarter-cell positions in Bacillus subtilis (41) and in E. coli (36) at least raises the possibility that plasmid duplication occurs at the mid-cell position and that after duplication the replication complexes with the attached plasmid clusters segregate to quarter-cell positions prior to the next round of cell division. Using a temperature-sensitive replication initiation protein for the F plasmid, evidence has been obtained in support of duplication of this plasmid at the mid-cell immediately followed by movement of each of the duplicated plasmids to their respective quarter-cell positions (51). Consistent with the notion that DNA replication proteins play a role in the segregation of plasmids, or plasmid clusters, to new fixed positions is the report of Miller and Cohen (46) that DNA replication proteins contribute to plasmid pSC101 segregation in addition to their role in replication of this plasmid.…”

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Gyrase Inhibitors and Thymine Starvation Disrupt the Normal Pattern of Plasmid RK2 Localization in Escherichia coli

2005

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Multicopy plasmids in Escherichia coli are not randomly distributed throughout the cell but exist as defined clusters that are localized at the mid-cell, or at the 1/4 and 3/4 cell length positions. To explore the factors that contribute to plasmid clustering and localization, E. coli cells carrying a plasmid RK2 derivative that can be tagged with a green fluorescent protein-LacI fusion protein were subjected to various conditions that interfere with plasmid superhelicity and/or DNA replication. The various treatments included thymine starvation and the addition of the gyrase inhibitors nalidixic acid and novobiocin. In each case, localization of plasmid clusters at the preferred positions was disrupted but the plasmids remained in clusters, suggesting that normal plasmid superhelicity and DNA synthesis in elongating cells are not required for the clustering of individual plasmid molecules. It was also observed that the inhibition of DNA replication by these treatments produced filaments in which the plasmid clusters were confined to one or two nucleoid bodies, which were located near the midline of the filament and were not evenly spaced throughout the filament, as is found in cells treated with cephalexin. Finally, the enhanced yellow fluorescent protein-RarA fusion protein was used to localize the replication complex in individual E. coli cells. Novobiocin and nalidixic acid treatment both resulted in rapid loss of RarA foci. Under these conditions the RK2 plasmid clusters were not disassembled, suggesting that a completely intact replication complex is not required for plasmid clustering.The development of methods for tagging bacterial plasmids with specific fluorescent fusion proteins or by fluorescence in situ hybridization has greatly facilitated the microscopic analysis of the location and movement of plasmids in a bacterial cell during growth and division. Contrary to the generally accepted view that plasmids randomly diffuse throughout the cell, it has now been shown that the low-copy-number plasmids F (16, 18, 49), P1 (16, 45), and R1 (31, 63) and the multicopy plasmids RK2 and pUC (4,25,53,54) are not randomly distributed throughout out an Escherichia coli cell but are present as clusters at preferred locations. Using differentially labeled probes and fluorescence in situ hybridization analysis, it has been shown that except for plasmid R1, which is located at mid-cell or at the cell poles (31), clusters of plasmids F, P1, RK2, and pUC generally are located at the mid-cell position in newborn E. coli cells and at the 1/4 and 3/4 positions in larger cells (16,25,44). For these plasmids it has been shown further that the movement of newly duplicated plasmid clusters from the mid-cell to the quarter-cell positions occurs with relatively rapid kinetics (16,18,25,44,49).Much has yet to be learned about cell-or plasmid-specified factors that are responsible for the localization and movement of plasmid clusters. It has been shown for plasmids F (18, 49), P1 (6, 44), R1 (63), R27 (39), and pB171 (5) that the...

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Behavior of Sister Copies of Mini-F Plasmid after Synchronized Plasmid Replication inEscherichia coliCells

Cited by 41 publications

References 18 publications

Subcellular Positioning of F Plasmid Mediated by Dynamic Localization of SopA and SopB

Subcellular Positioning of F Plasmid Mediated by Dynamic Localization of SopA and SopB

Multicopy Plasmids Affect Replisome Positioning inBacillus subtilis

Gyrase Inhibitors and Thymine Starvation Disrupt the Normal Pattern of Plasmid RK2 Localization in Escherichia coli

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