Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Bacterial DNA replication”

Search indexed PubMed citations on genomics, clinical trials, systematic reviews and public health. Explore titles, authors and supplied subject terms, then open the PubMed record.

Quote a phrase for an exact phrase match. Source license links do not imply unrestricted reuse.

At least 307 records · Page 17Linked to original sources
PubMed1976

Segregation of Bacillus subtilis chromosomes radioactively labeled during the first round of replication after germination of spores.

Spores of Bacillus subtilis W23 thy his were allowed to incorporate [3H]thymine for short periods of time either continuously from, or soon after, the start of the first round of replication after germination. They were then transferred to nonradioactive medium to allow growth into microcolonies (up to 12 cells), which were autoradiographed. The relative numbers of various types (major versus minor) of grain clusters associated with individual microcolonies throughout the populations were scored. Analysis of the results showed clearly that, in the majority of spores at least, only one chromosome was undergoing replication soon after the start of deoxyribonucleic acid synthesis. Furthermore, under the conditions used, no evidence for initiation of replication of a second chromosome within 25 min after the first could be obtained. Accepting that B. subtilis spores are essentially homogenous in deoxyribonucleic acid content, the results support the conclusion that the spore contains only one copy of the chromosome, not two.

Bacillus subtilis↗
PubMed2001

Telomere resolution in the Lyme disease spirochete.

The genus Borrelia includes the causative agents of Lyme disease and relapsing fever. An unusual feature of these bacteria is a genome that includes linear DNA molecules with covalently closed hairpin ends referred to as telomeres. We have investigated the mechanism by which the hairpin telomeres are processed during replication. A synthetic 140 bp sequence having the predicted structure of a replicated telomere was shown to function as a viable substrate for telomere resolution in vivo, and was sufficient to convert a circular replicon to a linear form. Our results suggest that the final step in the replication of linear Borrelia replicons is a site-specific DNA breakage and reunion event to regenerate covalently closed hairpin ends. The telomere substrate described here will be valuable both for in vivo manipulation of linear DNA in Borrelia and for in vitro studies to identify and characterize the telomere resolvase.

Borrelia burgdorferi↗
PubMed1988

The dnaK gene of Escherichia coli functions in initiation of chromosome replication.

A newly isolated dnaK mutant of Escherichia coli, which contains the mutation dnaK111, has been found to be conditionally defective in initiation of DNA replication. Mutant cells that were transferred to high temperature exhibited residual DNA synthesis before the synthesis stopped completely. Analysis of the DNA synthesized at high temperature by hybridization with probe DNAs for detection of DNA replicated in the origin (oriC) and terminal (terC) regions has revealed that this mutant is unable to initiate a new round of DNA replication at high temperature after termination of the round in progress. The cells exposed to high temperature were subsequently capable of initiating DNA replication at low temperature in a synchronous manner. DNA synthesis of this mutant became temperature resistant upon inactivation of the rnh gene, similar to that of dnaA mutants, although cell growth of the dnaK mutant with the inactive rnh gene remained temperature sensitive. The dnaK mutation prevented DNA synthesis of lambda bacteriophage at high temperature even in the absence of the rnh gene function.

Bacteriophage lambda↗
PubMed1994

The initiation cascade for chromosome replication in wild-type and Dam methyltransferase deficient Escherichia coli cells.

'Newborn' Escherichia coli B/r cells, obtained by membrane elution, were used to study the cell cycles of wild-type and Dam methyltransferase mutants. In wild-type cells, initiation of chromosome replication was synchronous and tightly controlled. In dam mutants, initiation was altered, but not random. We propose that this is due to the absence of an initiation cascade caused by liberated DnaA molecules, and that this cascade normally synchronizes initiation. The dam- cells contained mainly two, three or four replication origins, and this affected nucleoid partitioning as well as cell division. In cultures growing with a 50 min doubling time, a variety of cell cycles were present and half the origins were used every 25 min. Some cells had a 25 min interdivision time, whereas others had an interdivision time longer than the generation time. Partitioning of nucleoids containing unequal numbers of replication origins could also be readily observed by fluorescence microscopy in the dam mutant. Based upon these observations we propose that the dam mutant is also an initiation cascade mutant.

Bacterial Proteins↗
PubMed1984

Overinitiation of chromosome and plasmid replication in a dna Acos mutant of Escherichia coli K12. Evidence for dnaA-dnaB interactions.

The dnaAcos mutations are phenotypic suppressors of dnaAts46 that are co-transduced with dnaA, render the cell cold sensitive, and cause an excess of chromosome replication relative to cell mass when the cells are shifted from 42 degrees C to 32 degrees C. We have used pulse labelling and DNA-DNA hybridization to follow the effect of a temperature shift on the replication of the chromosome and of the plasmids pSC101, RTF-Tc, and lambda dv in such strains. After a shift of a dnaAcos strain from 42 degrees C to 32 degrees C (non-permissive temperature), initiation of the chromosome and replication of the plasmid pSC101 are stimulated, while the dnaA-independent plasmid RTF-Tc is not affected. The presence of pSC101 does not affect the level of overinitiation of the chromosome. The presence of lambda dv suppresses the cold sensitivity of dnaAcos mutants and allows the cells to grow at both 32 degrees C and 42 degrees C. The presence of lambda dv suppresses the overinitiation of chromosome and of pSC101 replication at 32 degrees C. Previous reports had shown that these suppressions involve an interaction between the dnaA product and the lambda P protein, which is also known to interact with dnaB. We show here that the mutant prophage P1 bac-crr, which produces high levels of a dnaB analogue, suppresses the dnaAcos phenotype, while wild type P1 does not. These results suggest that initiation involves interactions between the dnaA and dnaB products.

Chromosomes, Bacterial↗
PubMed1979

Selective association of the chromosome with membrane in a stable L-form of Bacillus subtilis.

A stalbe L-form (Sal-1) of Bacillus subtilis was found to have retained a markedly modified chromosome-membrane association when compared to intact cells. The membrane-deoxyribonucleic acid complex of the L-form was similar to that of its parental strain in quantity and stability. Genetic analysis of the L-form membrane-deoxyribonucleic acid complex revealed enrichment for markers close to the replication origin, but not for internal markers, indicating preferential attachment of the origin of chromosomal replication to the membrane. These results are in close agreement with those found for the parental bacterial form. In contrast, the replication termius region was not preferentially attached to the membrane of the L-form, even though it is enriched in the bacterial form. The association of the chromosome with the membrane at the replication terminus does not appear to be necessary for cell growth and separation, but because the L-form divides aberrantly, it may be one of the factors required for normal deoxyribonucleic acid segregation and septation.

Bacillus subtilis↗
PubMed1989

Sequence limits of DNA strands in the arrest replication fork at the Bacillus subtilis chromosome terminus.

The DNA sequence limits of the leading and lagging strands in the arrested clockwise replication fork at the terminus of the Bacillus subtilis chromosome have been investigated. On the basis of hybridization to synthetic oligonucleotides corresponding to known positions in the terminus region sequence it has been shown that neither the leading nor lagging strands, as they approach terC, traverse the distal inverted repeat, IRI. But a small fraction of the leading strands pass through the proximal inverted repeat, IRII. This is consistent with IRI being the functional inverted repeat in arresting the clockwise fork. But most of the forks appear to stop at least 100 nucleotides short of IRI, and at various positions extending over a distance of at least 100 nucleotides.

Bacillus subtilis↗
PubMed1998

High-affinity binding sites for the initiator protein DnaA on the chromosome of Escherichia coli.

The initiator protein DnaA of Escherichia coli binds with unusually high affinity to five regions on the chromosome, in addition to the replication origin, oriC. Using a solid-phase DNA binding assay, in which the DNA binding C-terminal domain of DnaA is bound via a biotin tag to magnetic beads, we could fish only fragments with these six regions from different chromosomal digests. Except for oriC, these fragments contain only one or two consensus DnaA binding sites, DnaA boxes. The distribution of these high-affinity DnaA boxes on the chromosome is random.

Bacterial Proteins↗
PubMed1975

Control of chromosome replication in thymine-requiring strains of Bacillus subtilis 168.

Study of the replication pattern of a number of B. subtilis 168 strains under controlled physiological conditions revealed great interstrain variation in control of replication. Replication patterns were calculated from ratios of purA16/leu-8 and purA16/metB5 transformation frequency. The thymine-independent strains are under strict regulation with an average of one replication position per chromosome during log phase. After starvation for required amino acids or sporulation, the chromosome is in a completed state with no replication forks (class I). In contrast, several thymine-requiring strains (class III) have an average of three to four replication positions per chromosome during log phase (multiforked replication) of which one to two remain uncompleted after amino acid starvation or sporulation. The other thymine-requiring strains studied are intermediate (class II) in that they have an average of two replication positions per chromosome during log phase and one after amino acid starvation or sporulation. Pulse chase experiments indicate that the deoxyribonucleic acid which is close to the chromosomal origin on each branch of the multiforked chromosome is bound to a rapidly sedimenting cellular fraction, presumably membrane.

Bacillus subtilis↗
PubMed1984

The dnaA protein complex with the E. coli chromosomal replication origin (oriC) and other DNA sites.

Specific binding of dnaA protein to the E. coli origin of replication (oriC) is essential for initiation of chromosomal replication. Based on binding of plasmid DNA fragments, dnaA protein recognizes in addition to oriC a number of specific sites: within or near the replication origins of pSC101, pBR322, and ColE1; within the regulatory regions of the dnaA and "X-protein" genes; and in IRL-Tn5. All strong binding sites share a common 9 bp sequence, 5'-TTATCACACAA, repeated at four conserved positions within oriC. As shown by DNAase I footprinting, dnaA protein binding is highly cooperative, covering 250 bp at oriC and 100 bp at single 9 bp sequence sites. Consistent with filter-binding and nuclease-protection studies, complexes of 20 to 30 dnaA monomers are visualized at oriC and other sites by electron microscopy. The functions of dnaA complexes and 9 bp sequences at these sites are discussed.

Bacterial Proteins↗
PubMed1992

RecA protein of Escherichia coli and chromosome partitioning.

Escherichia coli cells deficient in RecA protein frequently contain an abnormal number of chromosomes after completion of ongoing rounds of DNA replication. This suggests that RecA protein may be required for correct timing of initiation of DNA replication; however, we show here that initiation of DNA replication is properly timed in recA mutants. We also find that more than 10% of recA mutant cells contain no DNA. These anucleate cells appear to arise from partitioning of all the DNA into one daughter cell and no DNA into the other daughter cell. Based on these and previously published results, we propose that RecA protein is required for equal partitioning of chromosomes into the two daughter cells.

Cell Cycle↗
PubMed1990

DNA replication in Escherichia coli is initiated by membrane detachment of oriC. A model.

An adequate model for the initiation of chromosome replication in Escherichia coli should explain why the introduction of multiple copies of the chromosomal origin of replication, oriC, does not perturb cells seriously and why such multiple origins are replicated synchronously; it should explain why the key initiator protein, DnaA, is activated in vitro by binding specifically to acidic phospholipids and why the Dam methyltransferase is essential for the correct timing of initiation; it should explain why phospholipid synthesis and fluidity are necessary for initiation. In the detachment model, presented here, cyclical changes in the phospholipid composition of the cytoplasmic membrane activate initiator proteins such as DnaA protein and cause origins to detach; this detachment allows torsional stresses to open 13mer sequences in oriC; DnaA assists in the serial opening of these sequences and guides the entry of the helicase to form a pre-priming complex and trigger initiation; the greater affinity of hemi-methylated origin for membrane is re-interpreted as a mechanism for preventing re-initiation.

Cell Cycle↗
PubMed1998

Effects of chromosome underreplication on cell division in Escherichia coli.

The key processes of the bacterial cell cycle are controlled and coordinated to match cellular mass growth. We have studied the coordination between replication and cell division by using a temperature-controlled Escherichia coli intR1 strain. In this strain, the initiation time for chromosome replication can be displaced to later (underreplication) or earlier (overreplication) times in the cell cycle. We used underreplication conditions to study the response of cell division to a delayed initiation of replication. The bacteria were grown exponentially at 39 degreesC (normal DNA/mass ratio) and shifted to 38 and 37 degreesC. In the last two cases, new, stable, lower DNA/mass ratios were obtained. The rate of replication elongation was not affected under these conditions. At increasing degrees of underreplication, increasing proportions of the cells became elongated. Cell division took place in the middle in cells of normal size, whereas the longer cells divided at twice that size to produce one daughter cell of normal size and one three times as big. The elongated cells often produced one daughter cell lacking a chromosome; this was always the smallest daughter cells, and it was the size of a normal newborn cell. These results favor a model in which cell division takes place at only distinct cell sizes. Furthermore, the elongated cells had a lower probability of dividing than the cells of normal size, and they often contained more than two nucleoids. This suggests that for cell division to occur, not only must replication and nucleoid partitioning be completed, but also the DNA/mass ratio must be above a certain threshold value. Our data support the ideas that cell division has its own control system and that there is a checkpoint at which cell division may be abolished if previous key cell cycle processes have not run to completion.

Cell Cycle↗
PubMed1980

Chloramphenicol releases a block in initiation of chromosome replication in a dnaA strain of Escherichia coli K12.

DNA-DNA hybridisation experiments show that chloramphenicol induces a burst of initiation from the oriC region of a dnaA46 mutant of Escherichia coli at 36.5 degrees C but not from the isogenic dnaA+ strain. Following this stimulation of initiation is in parallel with the induced stimulation of RNA synthesis caused by chloramphenicol in the same strain. This is consistent with the hypothesis that the stimulation of initiation in the dnaA mutant is the result of the stimulation of the synthesis of an RNA species.

Chloramphenicol↗
PubMed1976

Isolation of a DNA-protein complex containing a single DNA fragment which is at or near the replication origin of the Bacillus subtilis chromosome.

A chromosomal fragment containing purA, the nearest marker from the replication origin of the Bacillus subtilis chromosome, was highly purified as a complex containing at least proteins and being solubilized easily during cell lysis. The complex had a markedly higher sedimentation rate (70-120S) than the bulk of the solubilized DNA (40S). The electron microscopic observation showed the complex to be an aggregate of several DNA molecules with a local structure containing amorphous materials which stained black and bushes of RNA. This confirmed biochemical evidences suggesting that the complex is an intermolecular aggregate of the purA-DNA-protein-RNA complex.

Bacillus subtilis↗
PubMed1978

Topological considerations in the theory of replication of DNA.

An obvious difficulty of the Watson-Crick model of DNA is that the intertwining of the strands would seem to hinder their separation during replication. The nature of the difficulty is here made precise and is called the alignment problem. It is shown that the swivelase theory, found in current textbooks and thought to overcome the difficulty, does not in fact do so. The various conceivable solutions of the alignment problem are considered and rejected, leading to the conclusion that chromosomal DNA is not double-helical. An alternative model of DNA is discussed. In addition a topological classification of DNA denaturation processes is given, and an alternative interpretation of the gel electrophoresis experiments on circular duplex DNA is suggested.

Chromosomes, Bacterial↗
PubMed1970

Sequence of genes replicated in Salmonella typhimurium as examined by transduction techniques.

5-Bromouracil (BU) was pulsed into the genome of synchronously growing cells of an F(-) strain of Salmonella typhimurium of LT2. BU-labeled genes were transduced with P22 phage to a series of recipient auxotrophs. When BU was incorporated early in the replication cycle, the transducing markers that had hybrid densities were those that lie between 9 and 12 o'clock on the genetic map. When BU was incorporated during the terminal period of the synchronous cycle, the transducing particles had hybrid densities for genes that lie from 1 to 8 o'clock clockwise. When phages were prepared on cells in which the middle period was BU-labeled, transducing particles with hybrid densities appeared for genes that lie in two separate regions: between 7 and 9 o'clock and between 12 and 2 o'clock. Analysis of the map sequences of the transduced BU genes, the relative frequency of transduction for each marker, and the time sequence of replication led to a hypothesis that the origin of replication is near the isoleucine-valine gene on the chromosome map. As for direction of replication, several models were considered, including the possibility that replication may proceed in both directions in the same chromosome. It was also found that the aroB, cysG, and strA genes are cotransduced and strA and aroC are also cotransduced. The relative order of the four genes was found to be aroB, cysG, strA, aroC, but the orientation in the circular map was not determined.

Amino Acids↗