Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “Replication Timing”

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 577 records · Page 32Linked to original sources
PubMed2004

Regulation of early events in chromosome replication.

Eukaryotic genomes are replicated from large numbers of replication origins distributed on multiple chromosomes. The activity of these origins must be coordinated so that the entire genome is efficiently and accurately replicated yet no region of the genome is ever replicated more than once. The past decade has seen significant advances in understanding how the initiation of DNA replication is regulated by key cell-cycle regulators, including the cyclin dependent kinases (CDKs) and the anaphase promoting complex/cyclosome (APC/C). The assembly of essential prereplicative complexes (pre-RCs) at origins only occurs when CDK activity is low and APC/C activity is high. Origin firing, however, can only occur when the APC/C is inactivated and CDKs become active. This two step mechanism ensures that no origin can fire more than once in a cell cycle. In all eukaryotes tested, CDKs can contribute to the inhibition of pre-RC assembly. This inhibition is characterised both by high degrees of redundancy and evolutionary plasticity. Geminin plays a crucial role in inhibiting licensing in metazoans and, like cyclins, is inactivated by the APC/C. Strategies involved in preventing re-replication in different organisms will be discussed.

Anaphase-Promoting Complex-Cyclosome↗
PubMed2006

DnaA couples DNA replication and the expression of two cell cycle master regulators.

Cell cycle progression in Caulobacter is driven by the master transcriptional regulators CtrA and GcrA. The cellular levels of CtrA and GcrA are temporally and spatially out-of-phase during the cell cycle, with CtrA repressing gcrA transcription and GcrA activating ctrA transcription. Here, we show that DnaA, a protein required for the initiation of DNA replication, also functions as a transcriptional activator of gcrA, which in turn activates multiple genes, notably those involved in chromosome replication and segregation. The cellular concentration of DnaA is cell cycle-controlled, peaking at the time of replication initiation and gcrA induction. Regulated proteolysis of GcrA contributes to the cell cycle variations in GcrA abundance. We propose that DnaA couples DNA replication initiation with the expression of the two oscillating regulators GcrA and CtrA and that the DnaA/GcrA/CtrA regulatory cascade drives the forward progression of the Caulobacter cell cycle.

Bacterial Proteins↗
PubMed2006

Vaccinia-virus-induced cellular contractility facilitates the subcellular localization of the viral replication sites.

Poxviruses, such as vaccinia virus (VV), replicate their DNA in endoplasmic-reticulum-enclosed cytoplasmic sites. Here, we compare the dynamics of the VV replication sites with those of the attenuated strain, modified VV Ankara (MVA). By live-cell imaging, small, early replication sites of both viruses undergo motility typical of microtubule (MT)-motor-mediated movement. Over time, growing replication sites of VV collect around the nucleus in a MT-dependent fashion, whereas those of MVA remain mostly scattered in the cytoplasm. Surprisingly, blocking the dynein function does not impair the perinuclear accumulation of large VV replication sites. Live-cell imaging demonstrates that in contrast to small replication sites, large sites do not display MT-motor-mediated motility. Instead, VV infection induces cellular contractility that facilitates the collection of growing replication sites around the nucleus. In a subset of cells (30-40%), this VV-induced contractility is alternated by phases of directed cell migration, suggesting that the two processes may be linked. The MVA-infected cells do not display contractility or cell migration, supporting the idea that these cellular activities facilitate the efficient accumulation of the VV replication sites around the nucleus. We propose that the recently described cytoskeletal rearrangements induced by VV are a prerequisite for the observed cell contractility and migration activities that apparently contribute to the organization of the complex cytoplasmic life cycle of VV.

Animals↗
PubMed1988

[Do phonological and semantic processings of Kanji finish at the same time?].

Two experiments were designed based on Neisser's visual searching paradigm to compare visual, phonological and semantic processing times of two-character compound Kanji, the Japanese logographic script. In Exp. 1, four undergraduates were asked to scan 10-word lists visually, phonologically or semantically according to the task required to find out a target which is either identical with or similar to the word previously presented at the trial. The results indicated that visual processing is quickest, and phonological and semantic processing finish at the same time. In Exp. 2, two- and four-syllable words were used as stimuli. Five undergraduates' results suggested that the length of syllable has effect on neither phonological nor semantic processing time, and replicated the results of Exp. 1 suggesting that phonological and semantic processing finish at the same time.

Adult↗
PubMedDate not supplied

Only oriC and its flanking region are recovered from the complex formed at the time of initiation of chromosome replication in Escherichia coli.

The oriC region of Escherichia coli constructs a specific complex to associate with the outer membrane fraction (oriC complex). The oriC complex was periodically formed before as well as in the short period after initiation of DNA replication. Using the DNA extracted from outer membrane fractions of the cells just after initiation as a probe, the whole E. coli genomic library was assayed by plaque hybridization. DNA regions that hybridized with the probe corresponded to about a 100-kb length of chromosome DNA which included oriC. In addition, clones located in a counterclockwise direction from oriC were more preferentially hybridized among these positive clones. Thus, we conclude that the oriC region is a unique locus of the chromosome that binds to the outer membrane at the time of initiation of chromosome replication.

Cell Membrane↗
PubMed1993

Origin activation and formation of single-strand TG1-3 tails occur sequentially in late S phase on a yeast linear plasmid.

In order to understand the mechanisms leading to the complete duplication of linear eukaryotic chromosomes, the temporal order of the events involved in replication of a 7.5-kb Saccharomyces cerevisiae linear plasmid called YLpFAT10 was determined. Two-dimensional agarose gel electrophoresis was used to map the position of the replication origin and the direction of replication fork movement through the plasmid. Replication began near the center of YLpFAT10 at the site in the 2 microns sequences that corresponds to the 2 microns origin of DNA replication. Replication forks proceeded bidirectionally from the origin to the ends of YLpFAT10. Thus, yeast telomeres do not themselves act as origins of DNA replication. The time of origin utilization on YLpFAT10 and on circular 2 microns DNA in the same cells was determined both by two-dimensional gel electrophoresis and by density transfer experiments. As expected, 2 microns DNA replicated in early S phase. However, replication of YLpFAT10 occurred in late S phase. Thus, the time of activation of the 2 microns origin depended upon its physical context. Density transfer experiments established that the acquisition of telomeric TG1-3 single-strand tails, a predicted intermediate in telomere replication, occurred immediately after the replication forks approached the ends of YLpFAT10. Thus, telomere replication may be the very last step in S phase.

Chromosomes, Fungal↗
PubMed2004

Macroscopic folding and replication of the homogeneously staining region in late S phase leads to the appearance of replication bands in mitotic chromosomes.

The chromosomal G/R bands are alternating domains differing in their nucleotide sequence biases. The bands are also related to the time of replication: pulse-labeling during S phase makes the replication sites as visible as replication bands that are close to the G/R bands in mitotic chromosomes. We previously showed that a plasmid bearing a mammalian replication origin efficiently generated a chromosomal homogeneously staining region (HSR). Here, we analyze the replication of this artificial HSR and show that it was replicated at the last stage of S phase. The HSR was composed of plasmid repeats only; nonetheless, we found that replication sites pulse-labeled during late S phase appeared as bands in the mitotic HSR and their number was dependent on the length of the HSR. Therefore, replication bands might not arise from sequence information per se. To understand the chronological order of appearance of replication sites, we performed a double pulse-chase experiment using IdU and CldU. Replication of the entire HSR required 100-120 minutes. During this period, the replicated sites appeared as bands at the first and last stages, but in between were apparently scattered along the entire HSR. An analysis of S-phase nuclei revealed that the replication started at the periphery of the globular HSR domain, followed by initiation in the internal domain. The replicated HSR appeared as a ring or a pair of extended spirals in late G2-phase nuclei. To account for these findings, we present a model in which the HSR is folded as a coiled-coil structure that is replicated from the outside to the inside in S phase nuclei.

Cell Line, Tumor↗
PubMed1998

Sex-specific selection on time to remate in Drosophila melanogaster.

Female Drosophila melanogaster were artificially selected for fast and slow time to remate (denoted 'high' and 'low' selection regimes, respectively). Both selection regimes and a control were replicated three times. Correlated responses to selection in females and in males were measured. A significant direct response to selection for time to remating was found in females from both selection regimes. Remating frequency of females showed a correlated response only in the females from the lines selected for faster time to remating. Time to first mating of virgin females showed no correlated response in either selection regime. No correlated response was found in males for time to remate, remating frequency or time to first mating of virgins, indicating that genetic correlations between the sexes do not influence the evolution of these traits in this population of D. melanogaster. There was no direct response to artificial selection for the ability of first males to deter females from remating. However, we found that the genotype of the first male to mate with a female could influence her time to remate; base stock males were better at deterring females from remating than were males from any of the selection lines. Copyright 1998 The Association for the Study of Animal Behaviour.

Journal Article↗
PubMed1988

Sequence of centromere separation: differential replication of pericentric heterochromatin in multicentric chromosomes.

The dicentric and multicentric chromosomes in L cells and a brain tumor cell line of mouse display only one site of kinetochore formation associated with the 'active' centromere. The accessory or 'inactive' centromeres show premature separation. These cell lines were treated with 10(-6) M 5-bromodeoxyuridine (BrdUrd) followed by anti-BrdUrd antibody to study the pattern of replication of pericentric heterochromatin flanking the active vs inactive centromeres. Regardless of its quantity, heterochromatin around the inactive centromere replicates earlier than that associated with the active centromere. There appears to be a relationship between the timing of separation of a centromere and the timing of replication of pericentric heterochromatin. The premature replication of heterochromatin associated with an inactive centromere may be responsible for its premature separation and, hence, inactivity.

Animals↗
PubMed2003

Replicating mesenchymal cells in the condyle and the glenoid fossa during mandibular forward positioning.

The purpose of this study was to identify and quantify the temporal sequence of replicating mesenchymal cells during natural growth and mandibular advancement in the condyle and the glenoid fossa. One hundred fifty 35-day-old female Sprague-Dawley rats were randomly divided into 10 experimental groups (10 rats each) and 10 control groups (5 rats each). The experimental groups were fitted with appliances that positioned the mandible forward. One hour before the rats were killed, bromodeoxyuridine (BrdU) was intravenously injected into them. Sections were cut and stained with anti-BrdU antibody to evaluate the number of replicating mesenchymal cells. Cellular uptake of BrdU was quantified with the Leica Qwin (Leica Microsystem Imaging Solutions, Cambridge, United Kingdom) system. The results showed that the numbers of replicating mesenchymal cells during natural growth were highest in the posterior region of the condyle and the anterior region of the glenoid fossa. In the experimental groups, the posterior region had the highest number of replicating cells for both the condyle and the glenoid fossa, with the condyle having 2 to 3 times more replicating cells than the glenoid fossa. The number of replicating mesenchymal cells, which is genetically controlled, influences the growth potential of the condyle and the glenoid fossa. Mandibular protrusion leads to an increase in the number of replicating cells in the temporomandibular joint. Individual variations in the response to growth modification therapy could be a result of the close correlation between mesenchymal cell numbers and growth.

Activator Appliances↗
PubMed1979

Conservative segregation of parental histones during replication in the presence of cycloheximide.

Long stretches of protein-free, nonbeaded DNA were observed electron microscopically in nuclear spreads prepared from cells that had replicated their DNA in the absence of protein synthesis. The amount of this DNA increased with increasing time of replication in the presence of cycloheximide and was greatly decreased when replication was inhibited with 1-beta-D-arabinofuranosylcytosine (cytosine arabinoside). This DNA is considered to be "free" DNA because it has the same diameter as marker PM2 DNA and it is preferentially sensitive to DNase I digestion. Reversal of the cycloheximide block resulted in a burst of histone synthesis and repair of the depleted chromatin within 5 min. In addition, 26 presumptive replication forks were observed with beaded chromatin on two arms and free DNA on the third. These results suggest that new histones are usually deposited onto new DNA, that the cellular histone pool is very small, that histone migration is minimal in vivo for at least 18 hr, that for most fibers nuclesome assembly and segregation is conservative for stretches of DNA as long as 100 kbases, and that some part of the octameric histone core may remain bound to DNA during the replication process. The regularity we have observed for the assembly and segregation of nucleosomes is likely to be important for our understanding of how chromosomal information is segregated during development.

Cell Line↗
PubMed1978

Regulation of ribosomal RNA synthesis in Tetrahymena pyriformis.

Ribosomal RNA is synthesized at constant rate during most of the cell cycle in heat-shock synchronized populations of Tetrahymena pyriformis. Early in each macronuclear S-period the rate of synthesis increases abruptly, concomitant with replication of the genes coding for ribosomal RNA. The increase is prevented by inhibitors of DNA replication, added prior to the S-period. Similarly, in cultures synchronized by starvation/refeeding, inhibition of DNA replication, at the time when the rDNA is replicated, will prevent the normal increase in rate of RNA synthesis which follows refeeding. We conclude that inhibition of rDNA replication interferes with the synthesis of rRNA, and we suggest that with respect to rRNA synthesis a gene dosis effect is operating in fast-growing Tetrahymena cells.

Animals↗
PubMed2001

Changes on Schistosoma mansoni (Digenea: Schistosomatidae) worm load in Nectomys squamipes (Rodentia: Sigmodontinae) concurrently infected with Echinostoma paraensei (Digenea: Echinostomatidae).

The water rat, Nectomys squamipes, closely involved in schistosomiasis transmission in Brazil, has been found naturally infected simultaneously by Schistosoma mansoni and Echinostoma paraensei. Laboratory experiments were conducted to verify parasitic interaction in concurrent infection. It was replicated four times with a total of 42 water rats and essayed two times with 90 mice pre-infected with E. paraensei. Rodents were divided into three groups in each replication. A wild strain recently isolated from Sumidouro, RJ, and a laboratory strain of S. mansoni from Belo Horizonte (BH) was used. Rats infected with E. paraensei were challenged 4 weeks later with S. mansoni and mice 2 or 6 weeks after the infection with S. mansoni. Necropsy took place 8 weeks following S. mansoni infection. The N. squamipes treatment groups challenged with S. mansoni RJ strain showed a significant decrease (80 and 65%) in the S. mansoni parasite load when compared with their respective control groups. There was a significant change or no change in the hosts challenged with the BH strain. The persistence time of E. paraensei within host was extended in relation to control groups, with a consequent enhancement of the number of recovered worm. An E. paraensei strain-specific influence on S. mansoni parasitism is reported. This paper presents some experimental data about this interaction in N. squamipes and Mus musculus.

Animals↗
PubMed1987

Removal of serine phosphates from simian virus 40 large T antigen increases its ability to stimulate DNA replication in vitro but has no effect on ATPase and DNA binding.

The effect of phosphorylation on the ability of simian virus 40 large T antigen to stimulate DNA synthesis in vitro was tested. Treatment of affinity-purified large T antigen with calf intestinal alkaline phosphatase resulted in the removal of 70 to 80% of the phosphate residues. Only serine-bound phosphate residues were affected. Phosphatase-treated large T antigen stimulated in vitro DNA synthesis fourfold over the untreated control. The stimulation was strongest at early times of DNA replication. At later times, DNA replication proceeded at equal rates with dephosphorylated and untreated large T antigen. The ATPase activity of large T antigen was not affected by phosphatase treatment. The origin-binding activity of large T antigen was tested over a wide range of large T antigen to DNA ratios, including DNA excess, and in the presence and absence of carrier DNA. Under no condition was an effect of dephosphorylation of large T antigen on its DNA-binding activity observed. These findings might indicate that phosphorylation at serine residues modulates the interaction of large T antigen with cellular factors. During DNA synthesis large T antigen was substantially rephosphorylated by kinases in the HeLa cell extract. As shown by two-dimensional peptide mapping, this phosphorylation occurred at all known in vivo sites. No phosphatase and protease activities were detectable in the HeLa cell extract.

Adenosine Triphosphatases↗
PubMed2001

Surface cues reduce the latency to name rotated images of objects.

Jolicoeur (1985, Memory & Cognition 13 289-303) found a linear increase in the latency to name line drawings of objects rotated (0 degrees to 120 degrees) from the upright (0 degrees) in the initial trial block. This effect was much shallower in later blocks. He proposed that the initial effect may indicate that mental rotation is the default process for recognising rotated objects, and that the decrease in this effect, seen with practice, may reflect the increased use of learned orientation-invariant features. Initially, we were interested in whether object-colour associations that may be learned during the initial block, could account for the reduced latency to name rotated objects, seen in later blocks. In experiment 1 we used full-cue colour images of objects that depicted colour and other surface cues. Surprisingly, given that Jolicoeur's findings were replicated several times with line drawings, we found that even the initial linear trend in naming latency was shallow. We replicated this result in follow-up experiments. In contrast, when we used less-realistic depictions of the same objects that had fewer visual cues (ie line drawings, coloured drawings, greyscale images), the results were comparable to those of Jolicoeur. Also, the initial linear trends were steeper for these depictions than for full-cue colour images. The results suggest that, when multiple surface cues are available in the image, mental rotation may not be the default recognition process.

Adult↗
PubMed1997

Nuclear sites of herpes simplex virus type 1 DNA replication and transcription colocalize at early times postinfection and are largely distinct from RNA processing factors.

We have visualized the intracellular localization of herpes simplex virus (HSV) type 1 replication and transcription sites in infected HeLa cells by using direct labelling methods. The number of viral transcription foci increases in a limited way; however, the number of replication sites increases in a near-exponential manner throughout infection, and both replication and transcription sites are found buried throughout the nuclear interior. Simultaneous visualization of viral transcription and replication foci shows that the two processes colocalize at early times, but at later times postinfection, there are additional sites committed solely to replication. This contrasts with the situation in adenovirus-infected cells in which, throughout replication, sites of transcription are adjacent to but do not colocalize with sites of viral DNA replication. The data for an increase in HSV transcription sites suggest an initial phase of replication of input genomes which are then transcribed. Sites of HSV replication colocalize with viral DNA replication and packaging proteins but are largely distinct from the punctate distribution of small nuclear ribonucleoprotein particles. Very high multiplicities of infection have shown an upper limit of some 18 viral transcription foci per nucleus, suggesting cellular constraints on transcription site formation. Use of virus replication mutants confirms that the labelled foci are sites of viral RNA and DNA synthesis; in the absence of viral DNA replication functions, no replication foci and only a limited number of transcription foci were present. Absence of a packaging function had no apparent effect on transcription or replication site formation, illustrating that DNA packaging is not a prerequisite for ongoing DNA synthesis. Further, the essential HSV protein IE63 is required for efficient replication site formation at later times postinfection but is not required for transcription foci formation.

Cell Nucleus↗
PubMed1996

Coordinating DNA replication initiation with cell growth: differential roles for DnaA and SeqA proteins.

We describe here the development of a new approach to the analysis of Escherichia coli replication control. Cells were grown at low growth rates, in which case the bacterial cell cycle approximates that of eukaryotic cells with G1, S, and G2 phases: cell division is followed sequentially by a gap period without DNA replication, replication of the single chromosome, another gap period, and finally the next cell division. Flow cytometry of such slowly growing cells reveals the timing of replication initiation as a function of cell mass. The data show that initiation is normally coupled to cell physiology extremely tightly: the distribution of individual cell masses at the time of initiation in wild-type cells is very narrow, with a coefficient of variation of less than 9%. Furthermore, a comparison between wild-type and seqA mutant cells shows that initiation occurs at a 10-20% lower mass in the seqA mutant, providing direct evidence that SeqA is a bona fide negative regulator of replication initiation. In dnaA (Ts) mutants the opposite is found: the mass at initiation is dramatically increased and the variability in cell mass at initiation is much higher than that for wild-type cells. In contrast to wild-type and dnaA(Ts) cells, seqA mutant cells frequently go through two initiation events per cell division cycle, and all the origins present in each cell are not initiated in synchrony. The implications for the complex interplay amongst growth, cell division, and DNA replication are discussed.

Bacterial Outer Membrane Proteins↗
PubMed1975

S-phase dependent forms of DNA - nuclear membrane complexes in HeLa cells.

DNA - nuclear membrane complexes were isolated from HeLa cells and examined by either zone sedimentation analysis or isopycnic centrifugation in sucrose/CsCl gradients. The data suggest that the complexes formed during the first 10 min of the S-phase remain as stable structures throughout the cell cycle. Other DNA - nuclear membrane complexes are formed at later times during replication. These later complexes appear as multiple species and the association of DNA and the nuclear membrane seems to be of a transient nature. Together, these results suggest that both the replicative origins and the replication points of the DNA are associated with the nuclear membrane. Although the complexes formed at the start of the S-phase and at later times during the S-phase appear to differ, these differences may provide them with the needed properties to serve as spatial organizers for the temporal regulation of DNA replication.

Binding Sites↗