共查询到20条相似文献,搜索用时 31 毫秒
1.
Changes in chromatin structure during the mitotic cycle 总被引:3,自引:0,他引:3
P. W. Barlow 《Protoplasma》1977,91(2):207-211
Summary Optical density profiles of Feulgen-stained nuclei ofBryonia dioica at different stages of the mitotic cycle were determined. Nuclei in the G2 phase have a greater fraction of dense chromatin than nuclei in G1 phase. However, nuclei at the end of the S phase have dispersed chromatin of minimal density. Thus, chromatin density oscillates during the mitotic cycle of this species, consequently the progressive increase in density previously recorded throughout the intermitotic period of two other species (onion and mouse) cannot be a general rule. 相似文献
2.
Mouse two-celled embryos and blastulae were Feulgen stained and the DNA content of their nuclei was measured with an integrating microdensitometer. The cells considered on the basis of their nuclear DNA content to be in G1, S, and G2 phases of the cell cycle were selected and their total chromatin area and chromatin areas at different gray levels were measured by the image analyzing computer, Quantimet. The measurements were aimed at quantitation of several features of the chromatin morphology of cells in different functional states. The total area of chromatin was found to increase, and the mean density of chromatin to decrease, from the G1 to the G2 phase of the cell cycle in both two-celled embryos and blastulae. The area of chromatin decreased, and the mean density of chromatin increased, as embryos developed from two-celled to blastula stage. It was concluded that nuclear morphology in preimplantation mouse embryos depends on both the phase of the cell cycle and the stage of development. The method of image analysis described was found to be useful for quantitation of changes in chromatin morphology. 相似文献
3.
Changes in the phosphorylation of non-histone chromatin proteins during the cell cycle of HeLa S 3 cells 总被引:2,自引:0,他引:2
R D Platz G S Stein L J Kleinsmith 《Biochemical and biophysical research communications》1973,51(3):735-740
The phosphorylation of non-histone chromatin proteins in synchronized HeLa S3 cells was studied in 5 phases of the cell cycle: mitosis, G1, early and late S, and G2. The rate of non-histone chromatin protein phosphorylation was found to be maximal during G1 and G2, somewhat decreased during S phase, and almost 90% depressed during mitosis. Analysis of the phosphorylated non-histone chromatin proteins by SDS-acrylamide gel electrophoresis showed a heterogeneous pattern of phosphorylation as measured by labeling with 32P. Significant variations in the labeling pattern were seen during different stages of the cell cycle, and particular unique species appeared to be phosphorylated selectively during certain stages of the cycle. 相似文献
4.
CHROMATIN STRUCTURE AND THE CELL DIVISION CYCLE : Actinomycin Binding in Synchronized HeLa Cells 总被引:8,自引:4,他引:4
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Measurements of actinomycin-3H binding in synchronized HeLa cells reveal that the binding capacity of chromatin decreases progressively during the S phase despite a doubling of nuclear DNA content, reaches a minimal level during G2 and mitosis, and then increases gradually throughout the subsequent G1 interval. Since this pattern was evident in experiments with living cells, ethanol-fixed cells, and isolated nuclei, but not with purified DNA, the actinomycin binding profile may reflect changes in the degree of association between DNA and chromosomal proteins at different stages of the cell cycle. 相似文献
5.
David A. Prentice Robert A. Tobey Lawrence R. Gurley 《Biochimica et Biophysica Acta (BBA) - Gene Structure and Expression》1983,741(3):288-296
The use of DNAase I as a probe of chromatin structure is frequently fraught with problems of irreproducibility. We have recently evaluated this procedure, documented the sources of the problems, and standardized the method for reproducible results (Prentice and Gurley (1983) Biochim. Biophys. Acta 740, 134–144). We have now used this probe to detect differences in chromatin structure between cells blocked (1) in G1 phase by isoleucine deprivation, or (2) in early S phase by sequential use of isoleucine deprivation followed by release into the presence of hydroxyurea. The cells blocked in G1 phase have easily-digestible chromatin, while cells blocked in early S phase have chromatin which is much more resistant to DNAase I. These differences were found to be the result of diffusible factors found in the cytoplasm and nuclei of G1- and S-phase cells, respectively. The G1 cells contained a cytoplasmic factor which modulates the chromatin structure of S-phase nuclei to a more easily digestible state, while cells blocked in S phase contain a nuclear factor which modulates the chromatin structure of G1 nuclei to a state more resistant to digestion. DNAase I is much more sensitive to these cell cycle-specific chromatin changes than is micrococcal nuclease. The results indicate that, under controlled conditions, DNAase I should be a valuable probe for detecting chromatin structural changes associated with cell cycle traverse, differentiation, development, hormone action and chemical toxicity. 相似文献
6.
The proliferation rate of mammalian cells is regulated normally in the G1 phase of the cell cycle. During this phase, it is convenient to assign positive and negative roles to the molecular programs that regulate the duration of G1 and the phase transition from G1 to S phase. Density-dependent inhibition of cellular proliferation results in an increase in the duration of G1. This form of regulation is due to both secreted factors and cell—cell contact. Serum is mitogenic to a variety of mammalian cell types. Because quiescent cells enter S phase as a result of serum addition to culture media, serum is usually regarded as a source of positive regulatory growth factors. We have measured the length of the G1, S and G2+ M phases of NIH 3T3 cells during exponential growth as a function of cell density and serum concentration. The G1 length increases during exponential growth as a function of density while S and G2+ M are relatively constant. Further, this increase in G1 phase time, or density mediated negative regulation, is inhibited by increasing serum concentration. This phenotype is saturable between 10% to 20% serum. Serum concentrations above 2.5% are able to increase the rate of cell cycling (decrease the G1 phase time) by inhibiting density dependent negative regulation of NIH 3T3. 相似文献
7.
Ioanna-Eleni Symeonidou Panagiotis Kotsantis Vassilis Roukos Maria-Anna Rapsomaniki Hernán E. Grecco Philippe Bastiaens Stavros Taraviras Zoi Lygerou 《The Journal of biological chemistry》2013,288(50):35852-35867
Once-per-cell cycle replication is regulated through the assembly onto chromatin of multisubunit protein complexes that license DNA for a further round of replication. Licensing consists of the loading of the hexameric MCM2–7 complex onto chromatin during G1 phase and is dependent on the licensing factor Cdt1. In vitro experiments have suggested a two-step binding mode for minichromosome maintenance (MCM) proteins, with transient initial interactions converted to stable chromatin loading. Here, we assess MCM loading in live human cells using an in vivo licensing assay on the basis of fluorescence recovery after photobleaching of GFP-tagged MCM protein subunits through the cell cycle. We show that, in telophase, MCM2 and MCM4 maintain transient interactions with chromatin, exhibiting kinetics similar to Cdt1. These are converted to stable interactions from early G1 phase. The immobile fraction of MCM2 and MCM4 increases during G1 phase, suggestive of reiterative licensing. In late G1 phase, a large fraction of MCM proteins are loaded onto chromatin, with maximal licensing observed just prior to S phase onset. Fluorescence loss in photobleaching experiments show subnuclear concentrations of MCM-chromatin interactions that differ as G1 phase progresses and do not colocalize with sites of DNA synthesis in S phase. 相似文献
8.
Zainy M. Banjara Robert C. Briggs Lubomir S. Hnilica Janet L. Stein Gary S. Stein 《Molecular and cellular biochemistry》1985,67(2):101-110
Summary Antisera to 0.35 M NaCl extracts and residues of S phase HeLa nuclei were reacted with electrophoretically separated proteins
from the nuclei or nuclear material of HeLa cells synchronized in G1, S, G2 or M phases of the cell cycle. Quantitative evaluation of the peroxidase-antiperoxidase stained nitrocellulose transfers
(Western blots) revealed significant changes in the quantities of nuclear non-histone proteins during the cell cycle. Immunochemical
staining of electrophoretically separated nuclear antigens permits their selective detection in minute quantities and in the
presence of many additional proteins. 相似文献
9.
Taizo Motomura 《Phycological Research》1996,44(1):11-17
The cell cycle (nuclear division cycle) of a multinucleate green alga, Boergesenia forbesii (Harvey) Feldmann was studied using microspectrophotometry and BrdU incorporation techniques. Mitosis was observed frequently 1-4 h after the beginning of the light period, on a 16:8 h LD cycle at 25°C. Mitotic nuclei formed discrete patches. Other nuclei remained in the G1 period. The DNA synthetic phase (S phase) was estimated to last about 12 h from microspectrophotometric study using aphidicolin inhibition just before the S phase and release from it. The G2 period was estimated to be about 2 h, because a labeled prophase nucleus could be detected when the samples were labeled with BrdU continuously over 3 h. The incorporation pattern of BrdU changed through the S phase nucleus. In early S phase, BrdU staining was detected as many dots in the entire nucleus, while in late S phase, it was detected as several discrete regions along the nuclear membrane. Almost all nuclei in B. forbesii were in the G1 stage after nuclear division, and the nuclei in several patches of the cell simultaneously initiated DNA synthesis. Once the nuclei entered into S phase, these nuclei continued into G2 and mitosis. In other words, the cell cycle regulation of entrance into S phase from G1 is an important factor in the growth and morphogenesis in B. forbesii. 相似文献
10.
Summary The arrangement of centromeres, cluster formation and association with the nucleolus and the nuclear membrane were characterized in human lymphocytes during the course of interphase in a cell-phase-dependent manner. We evaluated 3 893 cell nuclei categorized by five parameters. The centromeres were visualized by means of indirect immunofluorescent labeling with anti-centromere antibodies (ACA) contained in serum of patients with CREST syndrome. The cell nuclei were classified as G0, G1, S, G2, Gl1 and early S phase by comparing microscopically identified groups of cell nuclei with flow cytometric determination of cell cycle stage of synchronized and unsynchronized lymphocyte cell cultures. Based on a discrimination analysis, a program was devised that calculated the probability for any cell nucleus belonging to the G0, G1, S, G2, G1 and early S phase using only two microscopic parameters. Various characteristics were determined in the G0, S, and G2 stages. A transition stage to S phase within G1 was detected. This stage shows centromere arrangements not repeated in later cell cycles and which develop from the dissolution of centromere clusters in the periphery of the nucleus during G0 and G1. S phase exhibits various non-random centromere arrangements and associations of centromeres with the nucleolus. G1 and early S phase of the second cell cycle display no characteristic centromere arrangement. The duplication of centromeres in G2 is asynchronous in two phases. For all cell phases a test for random distribution of the centromeres in the cell nucleus was performed. There is a distinct tendency for centromeres to be in a peripheral position during Go and G1; this tendency becomes weaker in S phase. Although the visual impression is a seemingly random distribution of centromeres in G2 and G1 statistical analysis still demonstrates a significant deviation from random distribution in favor of a peripheral location. Only the early S phase of the second cell cycle shows no significant deviation from a random distribution. 相似文献
11.
Summary The relationships between cell kinetics and nuclear transformations in regeneration were investigated in the planarianPolycelis nigra by means of image analysis. A SAMBA 200 cell image processor was used to compute densitometric, textural and morphological parameters on Feulgen-stained nuclei in the blastema and near the cut 2–96 h after decapitation. On the basis of these parameters, the phase of the cell cycle (G1–G0, S, G2 and M) was identified and the variations in the percentage of cells in the various phases as well as the blastema cell number were computed against time after decapitation. It was demonstrated that the transection is followed by the sequential wasting of the M, G2, S and G1–G0 compartments. The depletion of a compartment was interpreted as being responsible for the subsequent recovery observed in the next one. The results show that cell proliferation at the section level is not sufficient to account for the increase of the blastema cell number during the first 48 h of regeneration, since the doubling time is about 12 h while the average cycle time is 48 h. It is thus suggested that G1–G0 cells migrate toward the section level, at least during the first 2 days of regeneration. Analysis of the nuclear profiles demonstrated that there are two different classes of G1–G0 cells: one corresponding to mature cells with a lot of condensed chromatin distributed in clumps within the nucleus, the other to immature cells with chromatin regularly distributed according to a rather homogeneous pattern. About one G1–G0 cell out of five is immature at the section level before decapitation while four cells out of five are immature as early as 8 h after the cut. This early inversion of the ratio between mature and immature cells argues in favour of an immigration of immature G1–G0 cells to the young blastema, where they are expected to accomplish only one cell cycle, and thus gives rise to mature cells. 相似文献
12.
Adam G. Evertts Amity L. Manning Xin Wang Nicholas J. Dyson Benjamin A. Garcia Hilary A. Coller 《Molecular biology of the cell》2013,24(19):3025-3037
The transition between proliferation and quiescence is frequently associated with changes in gene expression, extent of chromatin compaction, and histone modifications, but whether changes in chromatin state actually regulate cell cycle exit with quiescence is unclear. We find that primary human fibroblasts induced into quiescence exhibit tighter chromatin compaction. Mass spectrometry analysis of histone modifications reveals that H4K20me2 and H4K20me3 increase in quiescence and other histone modifications are present at similar levels in proliferating and quiescent cells. Analysis of cells in S, G2/M, and G1 phases shows that H4K20me1 increases after S phase and is converted to H4K20me2 and H4K20me3 in quiescence. Knockdown of the enzyme that creates H4K20me3 results in an increased fraction of cells in S phase, a defect in exiting the cell cycle, and decreased chromatin compaction. Overexpression of Suv4-20h1, the enzyme that creates H4K20me2 from H4K20me1, results in G2 arrest, consistent with a role for H4K20me1 in mitosis. The results suggest that the same lysine on H4K20 may, in its different methylation states, facilitate mitotic functions in M phase and promote chromatin compaction and cell cycle exit in quiescent cells. 相似文献
13.
SEPARATION OF NUCLEI REPRESENTING DIFFERENT PHASES OF THE GROWTH CYCLE FROM UNSYNCHRONIZED MAMMALIAN CELL CULTURES 总被引:5,自引:3,他引:2
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Nuclei have been isolated from unsynchronized cultures of Chinese hamster fibroblasts after varying intervals of growth following the incorporation of thymidine -3H for 20 min. These nuclei were fractionated by unit gravity sedimentation in a stabilizing density gradient of sucrose, and fractions were analyzed for the concentration of nuclei, DNA, and radioactivity. A more rapidly sedimenting population of nuclei in the G2 phase of the cell cycle was separated from a group of nuclei in the G1 phase, and nuclei in progressive stages of DNA synthesis (S phase) were distributed between these two regions. The fractionation of intact cells by sedimentation according to their position in the cell cycle was found to be less satisfactory than the corresponding separation of nuclei. This probably results from the continuous accumulation of mass within individual cells throughout the entire cell cycle, whereas most of the mass of a nucleus is replicated during a relatively narrow interval of the total cell cycle. 相似文献
14.
CELL CYCLE-SPECIFIC CHANGES IN HISTONE PHOSPHORYLATION ASSOCIATED WITH CELL PROLIFERATION AND CHROMOSOME CONDENSATION 总被引:13,自引:2,他引:11
下载免费PDF全文
![点击此处可从《The Journal of cell biology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Lawrence R. Gurley Ronald A. Walters Robert A. Tobey 《The Journal of cell biology》1974,60(2):356-364
Preparative polyacrylamide gel electrophoresis was used to examine histone phosphorylation in synchronized Chinese hamster cells (line CHO). Results showed that histone f1 phosphorylation, absent in G1-arrested and early G1-traversing cells, commences 2 h before entry of traversing cells into the S phase. It is concluded that f1 phosphorylation is one of the earliest biochemical events associated with conversion of nonproliferating cells to proliferating cells occurring on old f1 before synthesis of new f1 during the S phase. Results also showed that f3 and a subfraction of f1 were rapidly phosphorylated only during the time when cells were crossing the G2/M boundary and traversing prophase. Since these phosphorylation events do not occur in G1, S, or G2 and are reduced greatly in metaphase, it is concluded that these two specific phosphorylation events are involved with condensation of interphase chromatin into mitotic chromosomes. This conclusion is supported by loss of prelabeled 32PO4 from those specific histone fractions during transition of metaphase cells into interphase G1 cells. A model of the relationship of histone phosphorylation to the cell cycle is presented which suggests involvement of f1 phosphorylation in chromatin structural changes associated with a continuous interphase "chromosome cycle" which culminates at mitosis with an f3 and f1 phosphorylation-mediated chromosome condensation. 相似文献
15.
Cell cycle dependent chromosomal movement in pre-mitotic human T-lymphocyte nuclei 总被引:20,自引:0,他引:20
Fluorescent in situ hybridization with chromosome specific probes was used in conjunction with laser scanning confocal microscopy to assess the three-dimensional distribution of chromosomes in human T-lymphocyte nuclei. Cells in the G1-phase of the cell cycle exhibit a distinctly non-random chromosome organization: centromeric regions of the ten chromosomes examined are localized on the nuclear periphery, often making contact with the nuclear membrane, while telomeric domains are consistently localized within the interior 50% of the nuclear volume. Chromosome homolog pairing is not observed. Transition from the G1 to G2 cell cycle phase is accompanied by extensive chromosome movement, with centromeres assuming a more interior location. Chromosome condensation and chromatin depleted areas are observed in a small subset of G2 nuclei approaching mitosis. These results demonstrate that dynamic chromosome rearrangements occur in non-mitotic nuclei during the cell cycle.by L. Manuelidis 相似文献
16.
Quantitative and qualitative analyses were made of aberrations induced by 3 hitherto well-known mutagens, mitomycin C (MC), 5-bromodeoxyuridine (BUdR and hydroxylamine hydrocholride (HA), in muntjac chromosomes, during different stages of the cell cycle. The sensitivity ro MC was increased in G1, reached its maximum in early S and was considerably decreased in late S and G2 stage treated cells. BUdR induced maximal aberrations when given during the synthetic phase and the cells in G1 and G2 were least affected. The sensitivity of the cells to HA in terms of induced chromosomal aberrations increased as they moved through the cell cycle, i.e. more damage was observed in cells treated in late S and G2 stages than in those treated at G1 and early S stages. While there were defined patterns of cell-cylce stage-dependent sensitivity for all 3 chemicals, the chromosomal sites being preferentially affected by each were found to be specific and invariant at different stages. Thus, it is presumed that the functional state of such “preferred sites” at one or other stage of the cell cycle is the factor responsible for the stage-dependent sensitivity of a cell towards these chemicals. 相似文献
17.
Cell Cycle Dynamics of Cultured Coral Endosymbiotic Microalgae (Symbiodinium) Across Different Types (Species) Under Alternate Light and Temperature Conditions
下载免费PDF全文
![点击此处可从《The Journal of eukaryotic microbiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Lisa Fujise Matthew R. Nitschke Jörg C. Frommlet João Serôdio Stephen Woodcock Peter J. Ralph David J. Suggett 《The Journal of eukaryotic microbiology》2018,65(4):505-517
Dinoflagellates of the genus Symbiodinium live in symbiosis with many invertebrates, including reef‐building corals. Hosts maintain this symbiosis through continuous regulation of Symbiodinium cell density via expulsion and degradation (postmitotic) and/or constraining cell growth and division through manipulation of the symbiont cell cycle (premitotic). Importance of premitotic regulation is unknown since little data exists on cell cycles for the immense genetic diversity of Symbiodinium. We therefore examined cell cycle progression for several distinct SymbiodiniumITS2‐types (B1, C1, D1a). All types exhibited typical microalgal cell cycle progression, G1 phase through to S phase during the light period, and S phase to G2/M phase during the dark period. However, the proportion of cells in these phases differed between strains and reflected differences in growth rates. Undivided larger cells with 3n DNA content were observed especially in type D1a, which exhibited a distinct cell cycle pattern. We further compared cell cycle patterns under different growth light intensities and thermal regimes. Whilst light intensity did not affect cell cycle patterns, heat stress inhibited cell cycle progression and arrested all strains in G1 phase. We discuss the importance of understanding Symbiodinium functional diversity and how our findings apply to clarify stability of host‐Symbiodinium symbioses. 相似文献
18.
Summary Using flow cytometry, spherulating nuclei of Physarum isolated at the beginning of spherule wall formation were found to exhibit a DNA content corresponding to the G2 phase of the cell cycle, although 8% lower. Before the first mitosis after spherule germination, a very slight incorporation of 3H thymidine into DNA was observed that was too weak to correspond to S phase, strongly suggesting that nuclei are stopped in G2 phase inside the spherules. The lower value of nuclear DNA content found using flow cytometry of germinating spherules may not be related to DNA quantity, but may be due to a difference in chromatin organization during growth or spherulation, resulting in interference with the staining. 相似文献
19.
20.
Interaction of the Circadian Cycle with the Cell Cycle in Pyrocystis fusiformis 总被引:5,自引:3,他引:2
下载免费PDF全文
![点击此处可从《Plant physiology》网站下载免费的PDF全文](/ch/ext_images/free.gif)
Sweeney BM 《Plant physiology》1982,70(1):272-276
Dividing pairs or single cells of the large dinoflagellate, Pyrocystis fusiformis Murray, were isolated in capillary tubes and their morphology was observed over a number of days, either in a light-dark cycle or in constant darkness. Morphological stages were correlated with the first growth stage, G1, DNA synthesis, S, the second growth stage, G2, mitosis, M, and cytokinesis, C, segments of the cell division cycle. The S phase was identified by measuring the nuclear DNA content of cells of different morphologies by the fluorescence of 4′, 6-diamidino-2-phenylindole dichloride.
Cells changed from one morphological stage to the next only during the night phase of the circadian cycle, both under light-dark conditions and in continuous darkness. Cells in all segments of the cell division cycle displayed a circadian rhythm in bioluminescence. These findings are incompatible with a mechanism for circadian oscillations that invokes cycling in Gq, an hypothesized side loop from G1. All morphological stages, not only division, appear to be phased by the circadian clock.
相似文献