Effects of temperature and nutritional conditions on the mitotic cell cycle of Saccharomyces cerevisiae

1978 ◽  
Vol 31 (1) ◽  
pp. 71-78
Author(s):  
M.N. Jagadish ◽  
B.L. Carter
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Growth Rates ◽  
Mitotic Cell ◽  
S Phase ◽  
Yeast Cells ◽  
Batch Cultures ◽  
Nutritional Conditions ◽  
Limiting Nutrient ◽  
Effects Of Temperature

Yeast cells were cultivated at different growth rates in a chemostat by alterations in the flow of the limiting nutrient glucose and in batch cultures where variations in growth rate were achieved by alterations in the composition of nutrients. It was observed that the stage in the cycle at which S-phase was completed varied with growth rate. The faster the growth rate, the earlier the stage in the cycle in which completion of S-phase occurred. When stage in the cycle is converted into time before division it was observed that the time from completion of S-phase to cell division varied only slightly with growth rate except at extremely slow growth rates. Expansion of cell cycle transit time as the growth rate was slowed was achieved primarily by an expansion in time of the period from division to the completion of S-phase. In contrast, when cells were grown at different rates by alterations in the temperature of cultivation, completion of S-phase occurred at approximately the same stage in the cell cycle at all growth rates.

Regulation of the start of DNA replication in Schizosaccharomyces pombe

1999 ◽  
Vol 112 (6) ◽  
pp. 939-946 ◽  
Author(s):  
C.R. Carlson ◽  
B. Grallert ◽  
T. Stokke ◽  
E. Boye
Keyword(s):  
Cell Cycle ◽  
Flow Cytometry ◽  
Growth Rate ◽  
Schizosaccharomyces Pombe ◽  
Growth Rates ◽  
Cell Separation ◽  
Cell Mass ◽  
Nitrogen Sources ◽  
S Phase ◽  
G1 Phase

Cells of Schizosaccharomyces pombe were grown in minimal medium with different nitrogen sources under steady-state conditions, with doubling times ranging from 2.5 to 14 hours. Flow cytometry and fluorescence microscopy confirmed earlier findings that at rapid growth rates, the G1 phase was short and cell separation occurred at the end of S phase. For some nitrogen sources, the growth rate was greatly decreased, the G1 phase occupied 30–50% of the cell cycle, and cell separation occurred in early G1. In contrast, other nitrogen sources supported low growth rates without any significant increase in G1 duration. The method described allows manipulation of the length of G1 and the relative cell cycle position of S phase in wild-type cells. Cell mass was measured by flow cytometry as scattered light and as protein-associated fluorescence. The extensions of G1 were not related to cell mass at entry into S phase. Our data do not support the hypothesis that the cells must reach a certain fixed, critical mass before entry into S. We suggest that cell mass at the G1/S transition point is variable and determined by a set of molecular parameters. In the present experiments, these parameters were influenced by the different nitrogen sources in a way that was independent of the actual growth rate.

scholarly journals Coordination of Growth Rate, Cell Cycle, Stress Response, and Metabolic Activity in Yeast

2008 ◽  
Vol 19 (1) ◽  
pp. 352-367 ◽  
Author(s):  
Matthew J. Brauer ◽  
Curtis Huttenhower ◽  
Edoardo M. Airoldi ◽  
Rachel Rosenstein ◽  
John C. Matese ◽  
...  
Keyword(s):  
Gene Expression ◽  
Cell Cycle ◽  
Growth Rate ◽  
Stress Response ◽  
System Level ◽  
Cycle Phase ◽  
Instantaneous Growth Rate ◽  
Batch Cultures ◽  
Limiting Nutrient ◽  
Yeast Genes

We studied the relationship between growth rate and genome-wide gene expression, cell cycle progression, and glucose metabolism in 36 steady-state continuous cultures limited by one of six different nutrients (glucose, ammonium, sulfate, phosphate, uracil, or leucine). The expression of more than one quarter of all yeast genes is linearly correlated with growth rate, independent of the limiting nutrient. The subset of negatively growth-correlated genes is most enriched for peroxisomal functions, whereas positively correlated genes mainly encode ribosomal functions. Many (not all) genes associated with stress response are strongly correlated with growth rate, as are genes that are periodically expressed under conditions of metabolic cycling. We confirmed a linear relationship between growth rate and the fraction of the cell population in the G0/G1 cell cycle phase, independent of limiting nutrient. Cultures limited by auxotrophic requirements wasted excess glucose, whereas those limited on phosphate, sulfate, or ammonia did not; this phenomenon (reminiscent of the “Warburg effect” in cancer cells) was confirmed in batch cultures. Using an aggregate of gene expression values, we predict (in both continuous and batch cultures) an “instantaneous growth rate.” This concept is useful in interpreting the system-level connections among growth rate, metabolism, stress, and the cell cycle.

Ethylisopropylamiloride-sensitive pH control mechanisms modulate vascular smooth muscle cell growth

1991 ◽  
Vol 260 (3) ◽  
pp. C581-C588 ◽  
Author(s):  
A. Bobik ◽  
A. Grooms ◽  
P. J. Little ◽  
E. J. Cragoe ◽  
S. Grinpukel
Keyword(s):  
Cell Cycle ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Muscle Cells ◽  
Mitotic Cell ◽  
Ph Control ◽  
S Phase ◽  
Control Mechanisms ◽  
Aortic Smooth Muscle ◽  
Exchange Activity

The reported effects of alterations in Na-H exchange activity on mitogenesis are variable and appear dependent on the cell type examined. We examined the effects of reductions in ethylisopropylamiloride (EIPA)-sensitive pH-regulating mechanisms including Na-H exchange and alterations in intracellular pH (pHi) on the growth characteristics of rat aortic smooth muscle cells (RASM) cultured in serum-containing bicarbonate-buffered medium. Exposure of RASM replicating in bicarbonate-containing medium to the Na-H exchange inhibitors EIPA, dimethylamiloride (DMA), or amiloride (A) attenuated their replication rate. The order of potency of the inhibitors (EIPA greater than DMA much greater than A) was similar to their documented effects on Na-H exchange activity and to their order of potency for inhibiting recovery from CO2-induced acidosis in these cells. Reductions in pHi induced by lowering extracellular pH also attenuated the incorporation of [3H]-thymidine into DNA, while increases in pHi were associated with an acceleration in the rate of incorporation of [3H]thymidine into DNA. The effects of the Na-H exchange inhibitors on RASM replication were due to a reduction in the ability of the smooth muscle cells to enter the S phase of the mitotic cell cycle. This appeared predominantly the consequence of effects late within the G1 phase of the cell cycle. Concentrations of EIPA that markedly reduced the ability of RASM to enter S phase and to replicate also attenuated the increase in protein synthesis occurring 6-8 h after exposure to serum.(ABSTRACT TRUNCATED AT 250 WORDS)

scholarly journals CLN3 expression is sufficient to restore G1-to-S-phase progression in Saccharomyces cerevisiae mutants defective in translation initiation factor eIF4E

1999 ◽  
Vol 340 (1) ◽  
pp. 135-141 ◽  
Author(s):  
Parisa DANAIE ◽  
Michael ALTMANN ◽  
Michael N. HALL ◽  
Hans TRACHSEL ◽  
Stephen B. HELLIWELL
Keyword(s):  
Cell Cycle ◽  
S Phase ◽  
Translation Initiation Factor ◽  
Yeast Cells ◽  
Initiation Factor ◽  
G1 Phase ◽  
Mrna Levels ◽  
Wild Type ◽  
Phase Progression ◽  
Type Gene

The essential cap-binding protein (eIF4E) of Saccharomycescerevisiae is encoded by the CDC33 (wild-type) gene, originally isolated as a mutant, cdc33-1, which arrests growth in the G1 phase of the cell cycle at 37 °C. We show that other cdc33 mutants also arrest in G1. One of the first events required for G1-to-S-phase progression is the increased expression of cyclin 3. Constructs carrying the 5ʹ-untranslated region of CLN3 fused to lacZ exhibit weak reporter activity, which is significantly decreased in a cdc33-1 mutant, implying that CLN3 mRNA is an inefficiently translated mRNA that is sensitive to perturbations in the translation machinery. A cdc33-1 strain expressing either stable Cln3p (Cln3-1p) or a hybrid UBI4 5ʹ-CLN3 mRNA, whose translation displays decreased dependence on eIF4E, arrested randomly in the cell cycle. In these cells CLN2 mRNA levels remained high, indicating that Cln3p activity is maintained. Induction of a hybrid UBI4 5ʹ-CLN3 message in a cdc33-1 mutant previously arrested in G1 also caused entry into a new cell cycle. We conclude that eIF4E activity in the G1-phase is critical in allowing sufficient Cln3p activity to enable yeast cells to enter a new cell cycle.

Evaluation of genomic sequence-based growth rate methods for synchronized Synechococcus cultures

2021 ◽  
Author(s):  
Julia Carroll ◽  
Nicolas Van Oostende ◽  
Bess B. Ward
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Dna Replication ◽  
Growth Rates ◽  
Genomic Sequence ◽  
Niche Differentiation ◽  
Trophic Levels ◽  
Individual Species ◽  
Cell Counts

Standard methods for calculating microbial growth rates (μ) through the use of proxies, such as in situ fluorescence, cell cycle, or cell counts, are critical for determining the magnitude of the role bacteria play in marine carbon (C) and nitrogen (N) cycles. Taxon-specific growth rates in mixed assemblages would be useful for attributing biogeochemical processes to individual species and understanding niche differentiation among related clades, such as found in Synechococcus and Prochlorococcus . We tested three novel DNA sequencing-based methods (iRep, bPTR, and GRiD) for evaluating growth of light synchronized Synechococcus cultures under different light intensities and temperatures. In vivo fluorescence and cell cycle analysis were used to obtain standard estimates of growth rate for comparison with the sequence-based methods (SBM). None of the SBM values were correlated with growth rates calculated by standard techniques despite the fact that all three SBM were correlated with percentage of cells in S phase (DNA replication) over the diel cycle. Inaccuracy in determining the time of maximum DNA replication is unlikely to account entirely for the absence of relationship between SBM and growth rate, but the fact that most microbes in the surface ocean exhibit some degree of diel cyclicity is a caution for application of these methods. SBM correlate with DNA replication but cannot be interpreted quantitatively in terms of growth rate. Importance Small but abundant, cyanobacterial strains such as the photosynthetic Synechococcus spp. are essential because they contribute significantly to primary productivity in the ocean. These bacteria generate oxygen and provide biologically-available carbon, which is essential for organisms at higher trophic levels. The small size and diversity of natural microbial assemblages means that taxon-specific activities (e.g., growth rate) are difficult to obtain in the field. It has been suggested that sequence-based methods (SBM) may be able to solve this problem. We find, however, that SBM can detect DNA replication and are correlated with phases of the cell cycle but cannot be interpreted in terms of absolute growth rate for Synechococcus cultures growing under a day-night cycle, like that experienced in the ocean.

A murine replication protein accumulates temporarily in the heterochromatic regions of nuclei prior to initiation of DNA replication

1995 ◽  
Vol 108 (3) ◽  
pp. 927-934 ◽  
Author(s):  
M. Starborg ◽  
E. Brundell ◽  
K. Gell ◽  
C. Larsson ◽  
I. White ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Dna Replication ◽  
Mammalian Cells ◽  
Mitotic Cell ◽  
Yeast Cells ◽  
Metaphase Chromosomes ◽  
Licensing Factor ◽  
Mammalian Homologue ◽  
Initiation Of Dna Replication

We have analyzed the expression of the murine P1 gene, the mammalian homologue of the yeast MCM3 protein, during the mitotic cell cycle. The MCM3 protein has previously been shown to be of importance for initiation of DNA replication in Saccharomyces cerevisiae. We found that the murine P1 protein was present in the nuclei of mammalian cells throughout interphase of the cell cycle. This is in contrast to the MCM3 protein, which is located in the nuclei of yeast cells only between the M and the S phase of the cell cycle. Detailed analysis of the intranuclear localization of the P1 protein during the cell cycle revealed that it accumulates transiently in the heterochromatic regions towards the end of G1. The accumulation of the P1 protein in the heterochromatic regions prior to activation of DNA replication suggests that the mammalian P1 protein is also of importance for initiation of DNA replication. The MCM2-3.5 proteins have been suggested to represent yeast equivalents of a hypothetical replication licensing factor initially described in Xenopus. Our data support this model and indicate that the murine P1 protein could function as replication licensing factor. The chromosomal localization of the P1 gene was determined by fluorescence in situ hybridization to region 6p12 in human metaphase chromosomes.

Yeast Mutants That Produce a Novel Type of Ascus Containing Asci Instead of Spores

Genetics ◽  
1996 ◽  
Vol 144 (3) ◽  
pp. 979-989 ◽  
Author(s):  
Zhixiong Xue ◽  
Xiaoyin Shan ◽  
Alex Sinelnikov ◽  
Teri Melese
Keyword(s):  
Cell Cycle ◽  
Essential Gene ◽  
Mitotic Cell ◽  
Yeast Cells ◽  
Mitotic Cell Cycle ◽  
Spindle Formation ◽  
Bud Formation ◽  
Yeast Mutants ◽  
Two Hybrid

Abstract Tetraploid yeast cells lacking BFR1 or overexpressing an essential gene BBPl produce a novel type of ascus that contains asci instead of spores. We show here that the asci within an ascus likely arise because a/α spores undergo a second round of meiosis. Cells depleted of Bbplp or lacking Bfr1p are defective in a number of processes such as nuclear segregation, bud formation, cytokinesis and nuclear spindle formation. Furthermore, deletion of BFR1 or overexpression of BBP1 leads to an increase in cell ploidy, indicating that Bfr1p and Bbplp play roles in both the mitotic cell cycle and meiosis. Bfr1p and Bbp1p interact with each other in a two hybrid assay, further suggesting that they might form a complex important for cell cycle coordination.

scholarly journals Yeast Ppz1 protein phosphatase toxicity involves the alteration of multiple cellular targets

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Diego Velázquez ◽  
Marcel Albacar ◽  
Chunyi Zhang ◽  
Carlos Calafí ◽  
María López-Malo ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Transcription Factor ◽  
Protein Phosphatase ◽  
Mitotic Cell ◽  
Yeast Cells ◽  
Growth Defect ◽  
Cellular Targets ◽  
Major Mechanism ◽  
Bud Emergence ◽  
Phosphorylation Pattern

Abstract Control of the protein phosphorylation status is a major mechanism for regulation of cellular processes, and its alteration often lead to functional disorders. Ppz1, a protein phosphatase only found in fungi, is the most toxic protein when overexpressed in Saccharomyces cerevisiae. To investigate the molecular basis of this phenomenon, we carried out combined genome-wide transcriptomic and phosphoproteomic analyses. We have found that Ppz1 overexpression causes major changes in gene expression, affecting ~ 20% of the genome, together with oxidative stress and increase in total adenylate pools. Concurrently, we observe changes in the phosphorylation pattern of near 400 proteins (mainly dephosphorylated), including many proteins involved in mitotic cell cycle and bud emergence, rapid dephosphorylation of Snf1 and its downstream transcription factor Mig1, and phosphorylation of Hog1 and its downstream transcription factor Sko1. Deletion of HOG1 attenuates the growth defect of Ppz1-overexpressing cells, while that of SKO1 aggravates it. Our results demonstrate that Ppz1 overexpression has a widespread impact in the yeast cells and reveals new aspects of the regulation of the cell cycle.

scholarly journals Regulation of Cell Cycle Progression by Swe1p and Hog1p Following Hypertonic Stress

2001 ◽  
Vol 12 (1) ◽  
pp. 53-62 ◽  
Author(s):  
Matthew R. Alexander ◽  
Mike Tyers ◽  
Mireille Perret ◽  
B. Maureen Craig ◽  
Karen S. Fang ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Kinase Activity ◽  
Phosphorylation Site ◽  
S Phase ◽  
Mitogen Activated Protein Kinase ◽  
Yeast Cells ◽  
Hypertonic Stress ◽  
Cell Cycle Delay ◽  
G2 Phase ◽  
Morphogenesis Checkpoint

Exposure of yeast cells to an increase in external osmolarity induces a temporary growth arrest. Recovery from this stress is mediated by the accumulation of intracellular glycerol and the transcription of several stress response genes. Increased external osmolarity causes a transient accumulation of 1N and 2N cells and a concomitant depletion of S phase cells. Hypertonic stress triggers a cell cycle delay in G2 phase cells that appears distinct from the morphogenesis checkpoint, which operates in early S phase cells. Hypertonic stress causes a decrease in CLB2 mRNA, phosphorylation of Cdc28p, and inhibition of Clb2p-Cdc28p kinase activity, whereas Clb2 protein levels are unaffected. Like the morphogenesis checkpoint, the osmotic stress-induced G2 delay is dependent upon the kinase Swe1p, but is not tightly correlated with inhibition of Clb2p-Cdc28p kinase activity. Thus, deletion ofSWE1 does not prevent the hypertonic stress-induced inhibition of Clb2p-Cdc28p kinase activity. Mutation of the Swe1p phosphorylation site on Cdc28p (Y19) does not fully eliminate the Swe1p-dependent cell cycle delay, suggesting that Swe1p may have functions independent of Cdc28p phosphorylation. Conversely, deletion of the mitogen-activated protein kinase HOG1 does prevent Clb2p-Cdc28p inhibition by hypertonic stress, but does not block Cdc28p phosphorylation or alleviate the cell cycle delay. However, Hog1p does contribute to proper nuclear segregation after hypertonic stress in cells that lack Swe1p. These results suggest a hypertonic stress-induced cell cycle delay in G2 phase that is mediated in a novel way by Swe1p in cooperation with Hog1p.

scholarly journals Antagonistic regulation of Fus2p nuclear localization by pheromone signaling and the cell cycle

2009 ◽  
Vol 184 (3) ◽  
pp. 409-422 ◽  
Author(s):  
Casey A. Ydenberg ◽  
Mark D. Rose
Keyword(s):  
Cell Cycle ◽  
S Phase ◽  
Yeast Cells ◽  
Transcriptional Level ◽  
G1 Arrest ◽  
Cycle Arrest ◽  
Pheromone Signaling ◽  
Dependent Inhibition ◽  
Induced Proteins ◽  
Mating Projection

When yeast cells sense mating pheromone, they undergo a characteristic response involving changes in transcription, cell cycle arrest in early G1, and polarization along the pheromone gradient. Cells in G2/M respond to pheromone at the transcriptional level but do not polarize or mate until G1. Fus2p, a key regulator of cell fusion, localizes to the tip of the mating projection during pheromone-induced G1 arrest. Although Fus2p was expressed in G2/M cells after pheromone induction, it accumulated in the nucleus until after cell division. As cells arrested in G1, Fus2p was exported from the nucleus and localized to the nascent tip. Phosphorylation of Fus2p by Fus3p was required for Fus2p export; cyclin/Cdc28p-dependent inhibition of Fus3p during late G1 through S phase was sufficient to block exit. However, during G2/M, when Fus3p was activated by pheromone signaling, Cdc28p activity again blocked Fus2p export. Our results indicate a novel mechanism by which pheromone-induced proteins are regulated during the transition from mitosis to conjugation.

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