Search PubMed⌕ Search

SEARCH · Search PubMed

Results for “G2 Phase”

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 1,081 records · Page 60Linked to original sources
PubMed2003

Effect of BAP and IAA on the expression of G1 and G2 control points and G1-S and G2-M transitions in root meristem cells of Vicia faba.

Excised, carbohydrate-starved root meristems of Vicia faba subsp. minor have been used to investigate the impact of the auxin indole-3-acetic acid (IAA) and the cytokinin benzyl-6-aminopurine (BAP) on (1) the expression of Principal Control Points (PCPs) during the G1- and G2-phases of the cell cycle, and (2) the dynamics of sucrose-mediated resumption of DNA replication and mitosis (G1-to-S and G2-to-M transitions). Compared with the excised root tips starved in mineral medium without hormones, stationary phase meristems induced during continuous treatment with BAP, IAA, or a mixture of BAP+IAA, increased the number of G2 cells, producing characteristic profiles of nuclear DNA content. In medium containing 2% sucrose, BAP accelerated PCP1-->S and PCP2-->M, whereas continuous treatment with IAA resulted in marked prolongation of both transitions. Since the PCPs regulate progression through the key events of interphase and mitosis by interacting with cyclin dependent kinases (CDKs), these results seem to correspond with current data indicating functional connections between phytohormones, nutritional signals, gene expression and the cell division cycles in plants.

Benzyl Compounds↗
PubMed2005

Differential roles of XRCC2 in homologous recombinational repair of stalled replication forks.

Homologous recombination is an important mechanism in DNA replication to ensure faithful DNA synthesis and genomic stability. In this study, we investigated the role of XRCC2, a member of the RAD51 paralog family, in cellular recovery from replication arrest via homologous recombination. The protein expression of XRCC2, as well as its binding partner RAD51D, is dramatically increased in S- and G2-phases, suggesting that these proteins function during and after DNA synthesis. XRCC2 mutant irs1 cells exhibit hypersensitivity to hydroxyurea (HU) and are defective in the induction of RAD51 foci after HU treatment. In addition, the HU-induced chromatin association of RAD51 is deficient in irs1 mutant. Interestingly, irs1 cells are only slightly sensitive to thymidine and able to form intact RAD51 foci in S-phase cells arrested with thymidine. Irs1 cells showed increased level of chromatin-bound RAD51 as well as the wild type cells after thymidine treatment. Both HU and thymidine induce gamma-H2AX foci in arrested S-phase nuclei. These results suggest that XRCC2 is involved in repair of HU-induced damage, but not thymidine-induced damage, at the stalled replication forks. Our data suggest that there are at least two sub-pathways in homologous recombination, XRCC2-dependent and -independent, for repair of stalled replication forks and assembly of RAD51 foci following replication arrest in S-phase.

Animals↗
PubMed1997

Caffeine induces S-phase apoptosis in cis-diamminedichloroplatinum-treated cells, whereas cis-diamminedichloroplatinum induces a block in G2/M.

Caffeine overrides checkpoints in the G2 phase of the cell cycle by inhibiting DNA repair at this phase and increases the cytotoxicity of antitumor drugs, such as cis-diamminedichloroplatinum (CDDP). The enhanced cell death induced by caffeine is characterized by apoptosis. In this paper, we demonstrate that this apoptotic event occurs in S phase of the cell cycle, whereas CDDP induces a block in G2/M. DNA histogram analysis revealed that caffeine reduced G2 arrest in CDDP-treated EL-4 cells. In a synchronous population, the ratio of cyclin B:p34cdc2 was upregulated just before the cells went into the apoptotic pathway. A rapid increase in DNA fragmentation was detected at 12-24 h, when marked regression of G2/M phase was observed. Moreover, the degree of DNA fragmentation in CDDP + caffeine-treated cells was not reduced when the cell cycle was arrested at metaphase by exposure to the spindle-inhibitor nocodazole. It is possible that execution of the apoptotic program after treatment with caffeine did not require the EL-4 cells to reenter G1 phase. The apoptotic cell fraction in the group of CDDP + caffeine was recognized as an S population by bivariate analysis of apoptosis and DNA content. These results suggest that enhancement of the apoptotic activity of CDDP-treated cells by caffeine is not a G1-phase event but an S-phase-specific event, whereas cells were arrested in G2/M phase, and that it is regulated by G2 checkpoint-related proteins.

Animals↗
PubMed1995

Selectivity of cell cycle regulation of glucocorticoid receptor function.

The restricted expression of some genes to distinct stages of the cell cycle is often brought about through alterations in the activity and/or abundance of specific transcription factors. Many cells have been shown to be unresponsive to glucocorticoid hormone action during the G2 phase of the mammalian cell cycle, suggesting that some activities of the glucocorticoid receptor (GR), a ligand-activated transcription factor, are subjected to cell cycle control. We show here that GR insensitivity in G2 is selective, affecting receptor-mediated transactivation from a simple glucocorticoid response element, but not repression from a composite glucocorticoid response element. Since glucocorticoid-dependent down-regulation of GR protein levels is also unaffected in G2, distinct activities of the receptor that participate in this homologous down-regulation must be operating as effectively in G2-synchronized cells as in asynchronous cells. Finally, the phosphorylation state of the GR is altered in G2-synchronized cells reflecting, in part, both site-specific phosphorylation and dephosphorylation events. These results suggest that, while GR may be a target for cell cycle regulated kinases and phosphatases, the resulting changes in receptor phosphorylation have an impact only on selected GR functions.

Animals↗
PubMed1993

G2 delay induced by nitrogen mustard in human cells affects cyclin A/cdk2 and cyclin B1/cdc2-kinase complexes differently.

We investigated the temporal regulation of cyclin A- and B1-dependent kinases in human lymphoma cells treated with nitrogen mustard (HN2) and pentoxifylline, to determine whether the activity of these complexes correlated with cell cycle arrest induced by DNA damage. Cells were synchronized in G1/S, treated with HN2, and then postincubated with pentoxifylline. HN2-induced a protracted delay in G2 phase. This delay correlated with suppression of cyclin B1- and cdc2-kinase activities, and stabilization of hyperphosphorylated-cdc2 in the presence of similar cyclin B1 levels to those found in mitosis. HN2 had no discernible effect on the S phase activity of cyclin A- or cdk2-immune complexes. Entry of control cells into mitosis correlated with destruction of cyclin A, disappearance of cyclin A-bound cdk2 and decreased cdk2 kinase activity. G2 delay induced by HN2 was associated with stabilization of cyclin A, increased abundance of cyclin A-bound cdk2, and increased cdk2 activity. Cyclin A was also associated with cdc2, which, contrary to complexes containing cdk2, were only activated upon entry into mitosis. Pentoxifylline abrogated cell cycle arrest induced by aphidicolin and HN2 in human lymphoma cells. Pentoxifylline also reverted the activity of cyclin A- and B1-kinases in HN2-treated cells to approximately that observed in controls. Our findings suggest that delayed entry into mitosis following DNA damage correlates with suppression of cyclin B1/cdc2 and cyclin A/cdc2 complexes, while maintaining cyclin A/cdc2 complexes in an active state. Furthermore, we found that pentoxifylline disrupts the signal transduction pathway that regulates these complexes when damaged DNA is present, resulting in abrogation of cell cycle arrest.

Antigen-Antibody Complex↗
PubMed1995

Simplified and optimized kinetochore detection: cytogenetic marker for late-G2 cells.

Cytogenetic detection of kinetochore proteins using the CREST antibody coupled with secondary antibodies labeled with different fluorescent probes has been optimized for several in vitro mammalian cell lines. This study investigated selected parameters including the influence of common fixatives (methanol, ethanol, methanol:acetic acid (3:1)), detergents (Triton-X100, Tween), fluorescent probes (CY3, BODIPY, FITC), washing protocols, and an antifading agent (paraphenylenediamine) on the detection of kinetochore proteins in control and X-ray (240 kVp)-irradiated cells. Utilizing an optimized fixation and staining protocol, a brilliant visualization of kinetochores in interphase cells was obtained in control as well as X-ray-irradiated interhase cells. Application of this improved kinetochore staining methodology readily permits discriminating cells containing either single or paired kinetochores, the latter of which are characteristic of late-G2 phase and prophase cells.

Animals↗
PubMed2000

Recombination between homologous chromosomes does not play a dominant role in the formation of radiation-induced chromosomal aberrations.

PURPOSE: In mammalian cells, the relevance of homologous recombination in radiation-induced double-strand break (DSB) repair is not yet well understood. In the present work, the role of recombination between homologous chromosomes and homology-directed repair of DSB were studied, using X-ray-induced chromosomal aberrations as an end-point. MATERIALS AND METHODS: Human-hamster hybrid cells containing one or two copies of human chromosome 8 were used. If recombination between homologous chromosomes plays a dominant role in DSB repair, it is expected that X-irradiation of cells with two copies of chromosome 8 would result in a lower frequency of aberrations involving this chromosome compared with cells with only one copy of chromosome 8. The aberrations involving human chromosome 8 were detected by fluorescence in situ hybridization (FISH). Furthermore, a comparison between the hamster cell line XR-C1 (defective in non-homologous repair), CHO-9 (the wild-type cells) and the cell line XR-C1#8 (in which the defect of XR-C1 is complemented by human chromosome 8) was made to determine, indirectly, the involvement of homology-directed recombination in DSB repair. RESULTS: The observed frequencies of aberrations per human chromosome 8 were not significantly different between cells containing one or two copies of this chromosome. The frequency of chromatid-type aberrations was doubled in XR-C1 cells compared with CHO-9 and XR-C1#8 cells. CONCLUSIONS: In hamster cells, recombination between homologous chromosomes appears not to have a major role in the formation of radiation-induced chromosomal aberrations, while nonhomologous repair seems to be important in both the G and G2 phases of the cell cycle.

Animals↗
PubMed2005

[Studies on HIV/AIDS using the fission yeast Schizosaccharomyces pombe].

Human immunodeficiency virus (HIV) is a causative agent of acquired immunodeficiency syndrome (AIDS) and a member of Retrovirus family. The name of "retrovirus" is said to be derived from "reverse-transcribing oncogenic virus." Living up to its name, retrovirus has contributed to oncology, especially in the field of cancer pathogenesis. Retrovirus research has led to discovery of a number of oncogenes as well. Since the discovery of HIV in 1983, however, retrovirus has also been considered as an important etiologic agent that could incapacitate the cells involved in immune responses. As of the end of 2004, the number of people living with HIV/AIDS is estimated to be as large as 40 million. Every year, nearly 5 million people are newly infected with HIV, and 3 million people die of AIDS in the world, mainly in Africa and southeastern and southern Asia. Despite extensive studies, detailed mechanisms of HIV pathogenesis are still unclear, and efforts are being made to clarify functions of various HIV proteins and identify the cellular factors that could interact with the HIV proteins. One of the HIV accessory proteins, Vpr, causes the host cell cycle arrest at G2 phase, which may play an important pathogenic role in AIDS induction. Exploiting the fission yeast Schizosaccharomyces pombe useful for cell cycle studies, I've been trying to elucidate the mechanism by which Vpr induces the G2 arrest as presented in this review.

Acquired Immunodeficiency Syndrome↗
PubMedDate not supplied

Effect of cis-diammino-dichloro-platinum (II) on cell growth and cell cycle progression of a mouse ascites tumor growing in vivo.

Cell kinetic studies of cis-platin (CDDP) have been few with divergent results. In this study, the effect of 12.5 and 25 micrograms CDDP/animal corresponding to approximately 0.4 and 0.8 mg CDDP/kg body weight was described when treating the Bp8 ascites sarcoma growing in mice. From sequential studies of the total number of cells together with the composition of cells in the cell cycle, the flow of cells through the cell cycle was calculated. A dose-dependent cell growth inhibition was found with almost immediate effect following the higher dose, while an inhibitory effect on cell growth with the lower dose was delayed 24 hours. The cell flow from the various phases of the cell cycle for the first 24.48 hours following administration of the lower dose, was increased from the G1-phase and mitosis, with normal flow from the S-phase and depressed outflow from the G2-phase. Thereafter the flow rates from all cell cycle phases were decreased. For the higher dose, the outflow rates from all phases of the cell cycle were depressed up to 72 hours. The maximum platination of DNA was found one hour after treatment. The results indicate that the mechanisms responsible for the passage through the G2 stage of the cell cycle are more sensitive to the treatment of CDDP than replicative DNA synthesis. This experimental system may be used as a basis for further mechanistic studies of the antitumor effect of anticancer drugs.

Animals↗
PubMed1975

Effect of near ultraviolet and visible light on amoeba.

The near ultraviolet and visible light (VL) impinging at an intensity of 2-5 x 10(2) J s-1 m-2 for 2-5 h kills the mitotic and the early S-phase (0- to 15-min-old) amoebae. At the mid- and late S-period only a fraction of cells are killed by VL and G2 phase cells are quite resistant. Amoebae of all cell cycle stages show a delay in the first mitotic division. DNA synthesis, as measured by [3H]thymidine incorporation, is depressed in the VL-exposed early-S amoebae. A concurrent but temporary inhibition in [3H]leucine incorporation also occurs in these cells. However, no significant change in [3H]uridine incorporation has been found. To localize the site of lethal damage, nuclear transplantation studies were undertaken between the control amoebae and the amoebae treated with VL. The nucleus of a VL-exposed early S-phase cell recovers when transplanted immediately after VL exposure into an enucleate G2 cytoplasm but dies if grafted into an enucleat S-phase cytoplasm. The therapeutic effect of the G2 cytoplasm, although at a lower level, is also evident even when the treated early S-phase nucleus is implanted 20 h later, but not after 48 h, into the G2 cytoplasm. The amoeba cytoplasm shows resistance to VL-irradiation, can accept a control nucleus from any cell cycle stage, and function normally. The G2 nucleus also remains apparently unaffected to VL exposure and can survive when it is transfered to the control cytoplasm of any cell-cycle phase. All these findings are discussed in the light of the possible existence of a repair system against VL-induced damage in the G2-phase amoeba.

Amoeba↗
PubMed1997

Epithelial wound healing of the rat cornea after excimer laser ablation.

Rats with excimer corneal ablations (ArF 193 nm, 228 pulses, diameter 3,5 mm, 17.3 mJ per pulse, depth about 1/2 of corneal thickness) in one eye were killed 1, 2, 4, 6 and 13 days after treatment. The other eye served as control. The cell number per microscopic vision field, the labelling index (LI) and the mitotic rate (MR) were calculated for the peripheral, midperipheral and central areas of the corneal epithelium. The cell number showed a uniform depression in the remaining corneal epithelium at Day 1, normalizing from the centre to the periphery. The LI was only significantly increased at Day 1, while the MR was statistically significantly increased peripherally at Day 2 and in all areas at Day 6. However, when the corneal epithelium was evaluated as a whole, the MR was significantly increased at days 1, 2 and 6. The proliferative response of the epithelium was very homogenous irregardless of the distance to the original lesion. Both the migratory and the proliferative phases of the healing process seemed to be delayed when compared to the healing of pure epithelial wounds. However, the initiation of an increase in DNA synthesis seems not to be delayed, indicating that it is primarily the G2 phase that has been prolonged with this ablation procedure. The stromal thickness was increased from about one half of the normal values immediately after the treatment to above normal values at Day 4, thereafter decreasing to normal values at Day 13. Thus, the regenerative ability of the stroma is more pronounced in rats than in humans, but also in humans regeneration of the stroma can possibly explain the regression of the myopic shift seen some time after excimer laser treatment.

Animals↗
PubMed2006

Cohesin and DNA damage repair.

Replicated DNA molecules are physically connected by cohesin complexes from the time of their synthesis in S-phase until they are segregated during anaphase of the subsequent mitosis or meiosis. This sister chromatid cohesion is essential for the biorientation of chromosomes on the mitotic or meiotic spindle. In addition, cohesion is also essential during G2-phase of the cell cycle to allow repair of DNA double-strand breaks by homologous recombination. Although cohesion can normally only be established during S-phase, recent work in yeast has shown that DNA double-strand breaks induce the recruitment of cohesin to the damage site and lead to the de novo formation of cohesion at this site. It is unknown if similar mechanisms operate in higher eukaryotes, but in mammalian cells phosphorylation of the cohesin subunit Smc1 by the protein kinase Atm has been shown to be important for DNA repair. We discuss how cohesin and sister chromatid cohesion might facilitate the repair of damaged DNA.

Animals↗
PubMed1999

Involvement of G2-dependent DNA double-strand break repair in the formation of ultraviolet light B-induced chromosomal aberrations.

Wortmannin, an inhibitor of DNA double-strand break (DSB) repair added 19 h before harvest enhanced the incidence of ultraviolet light B (UVB)-induced chromatid aberrations in Chinese hamster V79 cells. Posttreatment with wortmannin for last 3 h of culture also enhanced the yield of breakage-type chromatid aberrations and suppressed the yield of exchange-type chromatid aberrations almost completely. Thus, the inhibition of DSB repair in the G2 phase stimulated the breakage-type aberration formation, while suppressing the exchange-type aberration formation. We propose the model of UVB-induced chromatid-type aberration formation which might be fully related to G2-dependent DSB repair pathway.

Androstadienes↗
PubMed2007

Skp2-mediated p27(Kip1) degradation during S/G2 phase progression of adipocyte hyperplasia.

p27(Kip1), an important regulator of Cdk2 activity and G1/S transition, is tightly regulated in a cell-type and condition-specific manner to integrate mitogenic and differentiation signals governing cell cycle progression. We show that p27 protein levels progressively declined from mid-G1 through late-G2 phase as density-arrested 3T3-L1 preadipocytes synchronously reentered the cell cycle during early stages of adipocyte differentiation. This dramatic fall in p27 protein accumulation was due, at least in part, to a decrease in protein stability. Specific inhibitors of the 26S proteasome were shown to completely block the decrease in p27 protein levels throughout G1, increase the abundance of ubiquitylated p27 protein, and inhibit G1/S transition resulting in G1 arrest. It is further demonstrated that p27 was phosphorylated on threonine 187 during S phase progression by Cdk2 and that phosphorylated p27 was polyubiquitylated and degraded. Furthermore, we demonstrate that Skp2 and Cks1 dramatically increased during S/G2 phase progression concomitantly with the maximal fall in p27 protein. Complete knockdown of Skp2 with RNA interference partially prevented p27 degradation equivalent to that observed with Cdk2 blockade suggesting that the SCF(Skp2) E3 ligase and other proteasome-dependent mechanisms contribute to p27 degradation during preadipocyte replication. Interestingly, Skp2-mediated p27 degradation was not essential for G1/S or S/G2 transition as preadipocytes shifted from quiescence to proliferation during adipocyte hyperplasia. Finally, evidence is presented suggesting that elevated p27 protein in the absence of Skp2 was neutralized by sequestration of p27 protein into Cyclin D1/Cdk4 complexes.

3T3-L1 Cells↗
PubMed2004

Tethering of HP1 proteins to chromatin is relieved by phosphoacetylation of histone H3.

Histone H3 lysine 9 methylation is associated with long-term transcriptional repression through recruitment of heterochromatin protein 1 (HP1) proteins. These proteins are believed to promote the formation of dense chromatin structures interfering with DNA accessibility. During the G2 phase of the cell cycle, HP1 proteins are delocalized from foci of pericentromeric heterochromatin, while a wave of H3 serine 10 phosphorylation is initiated within these regions. Here, we show that in vivo phosphorylation of serine 10 in G2 can occur on histone tails methylated on lysine 9. Unexpectedly, this modification favours rather than prevents HP1 binding to chromatin. Dissociation of HP1 from the methylated histone H3 tails is observed only after a third modification by acetylation of lysine 14, which occurs in prophase. We propose that phosphoacetylation of histone H3 could be a general mechanism allowing the cell to overcome HP1-mediated transcriptional repression.

Acetylation↗
PubMed1996

The REC1 gene of Ustilago maydis, which encodes a 3'-->5' exonuclease, couples DNA repair and completion of DNA synthesis to a mitotic checkpoint.

Mutation in the REC1 gene of Ustilago maydis results in extreme sensitivity to killing by ultraviolet light. The lethality of the rec1-1 mutant was found to be partially suppressed if irradiated cells were held artificially in G2-phase by addition of a microtubule inhibitor. This mutant was also found to be sensitive to killing when DNA synthesis was inhibited by external means through addition of hydroxyurea or by genetic control in a temperature-sensitive mutant strain defective in DNA synthesis. Flow cytometric analysis of exponentially growing cultures indicated that wild-type cells accumulated in G2 after UV irradiation, while rec1-1 cells appeared to exit from G2 and accumulate in G1/S. Analysis of mRNA levels in synchronized cells indicated that the REC1 gene is periodically expressed with the cell cycle and reaches maximal levels at G1/S. The results are interpreted to mean that a G2-M checkpoint is disabled in the rec1-1 mutant. It is proposed that the REC1 gene product functions in a surveillance system operating during S-phase and G2 to find and repair stretches of DNA with compromised integrity and to communicate with the cell cycle apparatus.

Base Sequence↗
PubMed1982

A comparison of mathematical methods for the analysis of DNA histograms obtained by flow cytometry.

Twelve methods for analysing FCM-histograms were compared using the same set of data. Some of the histograms that were analysed were simulated by computer and some were taken from experiments. Simulated data were generated assuming synchronously growing cell populations and (i) measurement coefficients of variation (CV) from 2 to 16%; (ii) constant measurement CV or VC's increasing from G1 to G2 phase, and (iii) varying fractions of cells in each phase. Simulated data were also generated assuming synchronous cell populations in which a block in early S phase was applied and released. DNA histograms were measured for L-929 cells at various times after mitotic selection. Labelling indices were also measured for these cells at the same time. The fractions of cells in the G1, S, and (G2 + M) phases were calculated by each analytical method and compared with the actual fractions used for simulation, or in case of experimental data, with autoradiographic results. Generally, all methods yielded reasonably accurate fractions of cells in each phase with relative errors in the range of 10-20%. However, most methods tended to overestimate G1 fractions and underestimate S fractions. In addition, variations in the shape of the S phase distribution often caused considerable errors. Phase fractions were also calculated for histograms of kinetically perturbed populations, simulated as well as experimental. The errors were only slightly larger than for histograms from asynchronously growing cell populations.

Analysis of Variance↗
PubMed1994

The effect of radiation on G2 blocks, cyclin B expression and cdc2 expression in human squamous carcinoma cell lines with different radiosensitivities.

The purpose of the present study was to investigate the role of cyclin B and cdc2 in the G2 delay and to test whether the magnitude of the G2 delay correlated with sensitivity to ionizing radiation in two human cell lines. Cell cycle delays were measured by flow cytometry after pulse labeling with bromodeoxyuridine, and expression of cell cycle control genes were measured in Western blots in radiosensitive SCC61 and radioresistant SQ20B cell lines. Flow cytometry data demonstrated that the duration of the G2 arrest was dose dependent in both cell lines, amounting to approximately 1.1 h/Gy. No difference was found between the cell lines in the length of the G2 block. Radiation exposure did not result in a decrease of cyclin B. Cyclin B protein levels in both asynchronous and synchronized populations in fact showed a dose dependent increase, concomitant with the rise in the fraction of cells in G2/M. Similarly, the cdc2 protein levels did not decrease after irradiation. However, it was found that the levels of hyperphosphorylated, and therefore inactive, kinase were significantly higher in irradiated cells than in unirradiated cells. The accumulation of this hyperphosphorylated form correlated with the arrest of cells in the G2 phase. Finally, immunocytochemical staining of cyclin B revealed an increase of this protein in the cytoplasm after irradiation and a decrease in nuclear staining. This differential localization could possibly account for the reduced nuclear phosphorylation of cdc2 kinase leading to the G2 arrest.

Blotting, Western↗