Cell Division In Response Research Articles

A delay in the Saccharomyces cerevisiae cell cycle that is induced by a dicentric chromosome and dependent upon mitotic checkpoints.

Dicentric chromosomes are genetically unstable and depress the rate of cell division in Saccharomyces cerevisiae. We have characterized the effects of a conditionally dicentric chromosome on the cell division cycle by using microscopy, flow cytometry, and an assay for histone H1 kinase activity. Activating the dicentric chromosome induced a delay in the cell cycle after DNA replication and before anaphase. The delay occurred in the absence of RAD9, a gene required to arrest cell division in response to DNA damage. The rate of dicentric chromosome loss, however, was elevated in the rad9 mutant. A mutation in BUB2, a gene required for arrest of cell division in response to loss of microtubule function, diminished the delay. Both RAD9 and BUB2 appear to be involved in the cellular response to a dicentric chromosome, since the conditionally dicentric rad9 bub2 double mutant was highly inviable. We conclude that a dicentric chromosome results in chromosome breakage and spindle aberrations prior to nuclear division that normally activate mitotic checkpoints, thereby delaying the onset of anaphase.

A delay in the Saccharomyces cerevisiae cell cycle that is induced by a dicentric chromosome and dependent upon mitotic checkpoints.

Dicentric chromosomes are genetically unstable and depress the rate of cell division in Saccharomyces cerevisiae. We have characterized the effects of a conditionally dicentric chromosome on the cell division cycle by using microscopy, flow cytometry, and an assay for histone H1 kinase activity. Activating the dicentric chromosome induced a delay in the cell cycle after DNA replication and before anaphase. The delay occurred in the absence of RAD9, a gene required to arrest cell division in response to DNA damage. The rate of dicentric chromosome loss, however, was elevated in the rad9 mutant. A mutation in BUB2, a gene required for arrest of cell division in response to loss of microtubule function, diminished the delay. Both RAD9 and BUB2 appear to be involved in the cellular response to a dicentric chromosome, since the conditionally dicentric rad9 bub2 double mutant was highly inviable. We conclude that a dicentric chromosome results in chromosome breakage and spindle aberrations prior to nuclear division that normally activate mitotic checkpoints, thereby delaying the onset of anaphase.

P34cdc2 homologue level, cell division, phytohormone responsiveness and cell differentiation in wheat leaves

Formation of a plant involves generation of new cells by the division cycle and development in these of specialised structure and metabolism. Specialisation is accompanied by a decreasing capacity for division, which declines with particular rapidity in cells of monocotyledonous plants such as the cereals. Here we report that in wheat leaves a homologue of the cell cycle control protein p34(cdc2) participates in the control of these developmental programmes. Accumulation of p34(cdc2) to a maximum level in dividing cells and the cessation of its accumulation during subsequent cell growth and expansion indicate that it contributes specifically to division. There is a decline in p34(cdc2) level as cell differentiation proceeds, in close parallel with the previously established decline of cell division in response to auxin hormones. A basal level of p34(cdc2) in fully differentiated cells that is one-sixteenth of that in dividing cells correlates with their loss of capacity to divide. We conclude that p34(cdc2) level is controlled in diverse multicellular eukaryotes and suggest that it is an important element in the switch from cell division to differentiation.

Cloning and sequence analysis of the Saccharomyces cerevisiae RAD9 gene and further evidence that its product is required for cell cycle arrest induced by DNA damage.

Procaryotic and eucaryotic cells possess mechanisms for arresting cell division in response to DNA damage. Eucaryotic cells arrest division in the G2 stage of the cell cycle, and various observations suggest that this arrest is necessary to ensure the completion of repair of damaged DNA before the entry of cells into mitosis. Here, we provide evidence that the Saccharomyces cerevisiae RAD9 gene, mutations of which confer sensitivity to DNA-damaging agents, is necessary for the cell cycle arrest phenomenon. Our studies with the rad9 delta mutation show that RAD9 plays a role in the cell cycle arrest of methyl methanesulfonate-treated cells and is absolutely required for the cell cycle arrest in the temperature-sensitive cdc9 mutant, which is defective in DNA ligase. At the restrictive temperature, cell cycle progression of cdc9 cells is blocked sometime after the DNA chain elongation step, whereas cdc9 rad9 delta cells do not arrest at this point and undergo one or two additional divisions. Upon transfer from the restrictive to the permissive temperature, a larger proportion of the cdc9 cells than of the cdc9 rad9 delta cells forms viable colonies, indicating that RAD9-mediated cell cycle arrest allows for proper ligation of DNA breaks before the entry of cells into mitosis. The rad9 delta mutation does not affect the frequency of spontaneous or UV-induced mutation and recombination, suggesting that RAD9 is not directly involved in mutagenic or recombinational repair processes. The RAD9 gene encodes a transcript of approximately 4.2 kilobases and a protein of 1,309 amino acids of Mr 148,412. We suggest that RAD9 may be involved in regulating the expression of genes required for the transition from G2 to mitosis.

Cloning and Sequence Analysis of the Saccharomyces cerevisiae RAD9 Gene and Further Evidence that Its Product Is Required for Cell Cycle Arrest Induced by DNA Damage

Procaryotic and eucaryotic cells possess mechanisms for arresting cell division in response to DNA damage. Eucaryotic cells arrest division in the G2 stage of the cell cycle, and various observations suggest that this arrest is necessary to ensure the completion of repair of damaged DNA before the entry of cells into mitosis. Here, we provide evidence that the Saccharomyces cerevisiae RAD9 gene, mutations of which confer sensitivity to DNA-damaging agents, is necessary for the cell cycle arrest phenomenon. Our studies with the rad9 delta mutation show that RAD9 plays a role in the cell cycle arrest of methyl methanesulfonate-treated cells and is absolutely required for the cell cycle arrest in the temperature-sensitive cdc9 mutant, which is defective in DNA ligase. At the restrictive temperature, cell cycle progression of cdc9 cells is blocked sometime after the DNA chain elongation step, whereas cdc9 rad9 delta cells do not arrest at this point and undergo one or two additional divisions. Upon transfer from the restrictive to the permissive temperature, a larger proportion of the cdc9 cells than of the cdc9 rad9 delta cells forms viable colonies, indicating that RAD9-mediated cell cycle arrest allows for proper ligation of DNA breaks before the entry of cells into mitosis. The rad9 delta mutation does not affect the frequency of spontaneous or UV-induced mutation and recombination, suggesting that RAD9 is not directly involved in mutagenic or recombinational repair processes. The RAD9 gene encodes a transcript of approximately 4.2 kilobases and a protein of 1,309 amino acids of Mr 148,412. We suggest that RAD9 may be involved in regulating the expression of genes required for the transition from G2 to mitosis.

A Non-Nodulating Alfalfa Mutant Displays Neither Root Hair Curling nor Early Cell Division in Response to Rhizobium meliloti

A Non-Nodulating Alfalfa Mutant Displays Neither Root Hair Curling nor Early Cell Division in Response to Rhizobium meliloti

A non-nodulating alfalfa mutant displays neither root hair curling nor early cell division in response to Rhizobium meliloti.

The early events in the alfalfa-Rhizobium meliloti symbiosis include deformation of epidermal root hairs and the approximately concurrent stimulation of cell dedifferentiation and cell division in the root inner cortex. These early steps have been studied previously by analysis of R. meliloti mutants. Bacterial strains mutated in nodABC, for example, fail to stimulate either root hair curling or cell division events in the plant host, whereas exopolysaccharide (exo) mutants of R. meliloti stimulate host cell division but the resulting nodules are uninfected. As a further approach to understanding early symbiotic interactions, we have investigated the phenotype of a non-nodulating alfalfa mutant, MnNC-1008 (NN) (referred to as MN-1008). Nodulating and non-nodulating plants were inoculated with wild-type R. meliloti and scored for root hair curling and cell divisions. MN-1008 was found to be defective in both responses. Mutant plants inoculated with Exo- bacteria also showed no cell division response. Therefore, the genetic function mutated in MN-1008 is required for both root hair curling and cell division, as is true for the R. meliloti nodABC genes. These observations support the model that the distinct cellular processes of root hair curling and cell division are triggered by related mechanisms or components, or are causally linked.

The influence of 17 beta-estradiol on patterns of cell division in the uterus.

Uteri from immature (21-day-old) and adult mice show different patterns of cell division in response to a physiological dose of 17 beta-estradiol. In the immature mouse uterus, estradiol increased stromal and epithelial cell proliferation by shortening the cell generation time (Tc). The epithelial Tc was reduced from 100 to 14.5 h; the stromal Tc was reduced to 16 h. In both cell types, the reduction in Tc was primarily due to a reduction of the G1 phase of the cell cycle. In the adult mouse uterus, estradiol stimulated epithelial but not stromal cell proliferation. Here, estradiol reduced the epithelial Tc from 86 to 13 h, mostly by reducing the G1 phase of the cell cycle. Therefore, estrogens stimulate uterine hyperplasia by selectively decreasing the G1 phase of the cell cycle in specific cell populations. At some point during reproductive tract maturation, uterine stromal cells lose their ability to divide in response to estrogen stimulation. A developmental study showed that in the intact mouse, the stromal cell population gradually lost its ability to divide in response to estrogen stimulation during days 22-52 after birth. In prepubertally ovariectomized mice, the stromal cell population showed a very low mitotic response to estrogen stimulation at all ages. Thus, an ovarian mechanism may regulate the change in stromal responsiveness to estrogen stimulation.

Yeast cells recover from mating pheromone alpha factor-induced division arrest by desensitization in the absence of alpha factor destruction.

Saccharomyces cerevisiae MATa cells arrest cell division in response to the mating pheromone, alpha factor. After some interval of time the cells resume division. It is demonstrated here that cells recover from division arrest by becoming insensitive to the alpha factor under conditions of low cell density (10(2) cells/ml) where no alpha factor destruction occurs. The time of desensitization occurs later at higher alpha factor concentrations. Using the technique of perfusion photomicroscopy developed in this study, it was found that 95% of the desensitized cells remain insensitive to the alpha factor for greater than or equal to 3 generations at the alpha factor concentration where recovery occurred. Upon recovery, cells have generation times which are similar or identical to the calculated time from the cdc28 "start" step of cell division to cell separation. Therefore, the abnormally large recovered cells behave as if they have no growth time requirement for cell division for several generations. Desensitization was found to occur asymmetrically for parent and daughter cells. While parents (cells which have budded) are insensitive to alpha factor, their daughters (cells which have never budded and which were formed from desensitized parents during continuous perfusion with alpha factor) show 54% with a delayed generation time compared to control daughter cells.

Isolation of cloned cDNAs to auxin-responsive poly(A) + RNAs of elongating soybean hypocotyl

Auxin-responsive cDNA clones have been isolated from a cDNA library prepared from elongating soybean hypocotyl poly(A)(+)RNA. The expression of two such sequences has been assessed by RNA blot hybridization analyses during normal developmental transitions in the soybean hypocotyl and during incubation of sections excised from the region of cell elongation. The concentrations of these poly(A)(+)RNAs are higher in the elongating zone than in the apical and mature zones of the hypocotyl. Both poly(A)(+)RNAs are depleted during incubation of the sections in the absence of auxin. The loss of one of these sequences (pJCW1) is prevented by the addition of auxin to the incubation medium while the other sequence (pJCW2) increases above the initial level in the presence of auxin. The addition of auxin to auxin-depleted tissue in which the sequences are depleted results in rapid accumulation of these poly(A)(+)RNAs; pJCW1 accumulates to the control level while pJCW2 increases well above the control level. These data along with others [Baulcombe, D. C. & Key, J. L. (1980) J. Biol. Chem. 255, 8907-8913] demonstrate directly a highly selective effect of auxin on the expression of a small number of mRNAs in tissues undergoing both cell elongation and cell division in response to auxin. Although the data are suggestive of a close association betwen auxin action and altered gene expression, a causal relationship is not established. It seems highly unlikely, however, that such specific effects of auxin on gene expression are unimportant in auxin physiology.

Hormonal control of uterine growth: the effect of hypothyroidism on estrogen-stimulated cell division.

The increase in mitotic indices of uterine luminal epithelium, stroma, and myometrium were determined as a function of time after the administration of a single dose of 17 beta-estradiol to euthyroid and hypothyroid rats. Hypothyroidism reduced the increase in the mitotic index 5-fold in the luminal epithelium, 6-fold in the stroma, and 9-fold in the myometrium. In addition to reducing mitotic indices, hypothyroidism also produced a shift of 12 h in the time course of estrogen-stimulated cell division of all uterine cell types relative to euthyroid animals. This shift in the time course of cell division was preceded by a shift in the time course of uterine DNA synthesis measured by tritiated thymidine incorporation. In contrast, hypothyroidism did not alter the magnitude or the time course of synthesis of 2-deoxyglucose-6-phosphate from 2-deoxyglucose after estrogenic stimulation. These results indicate that hypothyroidism decreases the ability of all major uterine cell types to undergo cell division in response to acute administration of estradiol, and also shifts the time course of the uterine growth response to the hormone.

Photoinduced division synchrony in permanently bleached Euglena gracilis

Ultraviolet light-induced, bleached Euglena clones exhibit synchronous steps of cell division in response to daily cycles of light and dark. The cyclic division activity, in the bleached cells, will persist in constant lighting conditions with a period, independent of temperature, of about 24 h. This persisting rhythm of cell division supports the hypothesis that this phase of the cell cycle may be coupled to the fluctuations of the endogenous circadian clock of the cell.Newly isolated bleached clones are sensitive to light in their growth rates and metabolic characteristics, showing light induced difference in substrate-stimulated respiration, and production of the polyglucan, paramylon. After repeated subculturing of a bleached clone the photosensitivity of the metabolic characteristics and of the growth rate are diminished along with the ability to photo-entrain division synchrony. Division control and the induction of cell synchrony in this organism apparently involve both the temporal influence of the endogenous cell clock and one or more other photosensitive reactions in the metabolism of the cell.A unique culture mixing technique utilizing the bleached Euglena, failed to support the hypothesis of the involvement of intercellular communication in the maintenance of cell synchrony in constant lighting conditions.