Pattern of end growth of the fission yeast Schizosaccharomyces pombe

1990 ◽  
Vol 36 (6) ◽  
pp. 390-394 ◽  
Author(s):  
Hisao Miyata ◽  
Machiko Miyata ◽  
Byron F. Johnson
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Schizosaccharomyces Pombe ◽  
Key Words ◽  
Fission Yeast ◽  
Doubling Time ◽  
Time Lapse ◽  
Critical Value ◽  
Wild Type ◽  
Time Lapse Photomicrography

The patterns of end growth of individual cells of Schizosaccharomyces pombe, wild-type cells (strain 972 h−), cells exposed to 8 mM hydroxyurea, and cdc mutants (cdc11-123 and cdc2-33), were investigated by time-lapse photomicrography. It was reconfirmed that there are three patterns of end growth: cells growing at the old end, at the new end, and at both ends from the beginning of the cell cycle. Cells that initiated growth at the old (new) end increased their growth rate at the new (old) end and became constant in their growth rate at the old (new) end when cells had their growth rate higher than a critical value: 0.08, 0.09, 0.08, and 0.11 μm/min in wild-type cells, cells exposed to hydroxyurea, cdc11-123 cells, and cdc2-33 cells, respectively. The critical value is proportional to the doubling time in length. Key words: extension, growth, fission yeast.

scholarly journals The effect of CO2 on the timing of cell cycle events in fission yeast Schizosaccharomyces pombe

1988 ◽  
Vol 89 (3) ◽  
pp. 433-439
Author(s):  
B. NOVÁK ◽  
J. HALBAUER ◽  
E. LÁSZLÓ
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Co2 Production ◽  
Complete Medium ◽  
Co2 Removal ◽  
Wild Type ◽  
In The Wild ◽  
G2 Phase

The effect of CO2 removal on the cell cycle phases of Schizosaccharomyces pombe has been examinedin minimal, aspartate-containing and complete medium. The removal of CO2 shortened the G2 phase of the cell cycle and arrested the cells in G1 phase in minimal medium. The G1 block caused by CO2 deprivation was demonstrated by transition-point and flow-cytometry analyses. The slow-down of anapleurotic CO2 fixation might be responsible for this effect, as aspartic acid could abolish the G1 block. The shortening of G2 phase in the wild-type cells was observed in every medium irrespective of whether the growth rate was changed or not. The experiments in which growth rate was not changed by CO2 shift-down suggest that this CO2 effect can be independent from its action on CO2-fixing steps in metabolism. Therefore we propose that CO2 inhibits mitosis infission yeast and we explain the proportionality between growth rate and cell size at mitosis found by Fantes & Nurse by this CO2 inhibition. The larger CO2 production in fast-growing cells leads to a higher CO2 concentration, which could exerta stronger inhibition of mitosis. A wee mutant, which has lost its mitotic size control, also shows the G1 block after CO2 deprivation, but its mitosis is insensitive to CO2. Comparing the respiration of wee and wild-type cells we conclude that CO2 inhibits the citric acid cycle in the wild type. The consequence of these results in the regulation of fission yeast cell cycle is discussed.

Patterns of extension growth of the fission yeast, Schizosaccharomyces pombe

1986 ◽  
Vol 32 (6) ◽  
pp. 528-530 ◽  
Author(s):  
H. Miyata ◽  
M. Miyata ◽  
Byron F. Johnson
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Dilution Rate ◽  
Progenitor Cells ◽  
Progenitor Cell ◽  
Chemostat Culture ◽  
Time Lapse ◽  
Extension Growth ◽  
Previous Cycle ◽  
Time Lapse Photomicrography

The growth of sausage-shaped cells of the fission yeast, Schizosaccharomyces pombe (strain NCYC 132), was followed in the second or third cycle by time-lapse photomicrography. Experimental cells were harvested from glucose-limited (0.2% glucose EMM3) chemostat culture (dilution rate, 0.125/h) and were plated onto a slide with EMM3 agar (2% glucose). By observing their extension patterns, we found some rules of extension growth. Thus, (1) all sibs with walls newly formed in the previous cycle, whose progenitor cells grew at the old end (followed Mitchison's rule), grow at the old end (also follow Mitchison's rule). (2) Sibs with old walls whose progenitor cell followed Mitchison's rule behave in one of three ways: (i) growth at the old end (follow Mitchison's rule); (ii) growth at the new end (violate Mitchison's rule); or (iii) growth at both ends (bipolar). (3) Both sibs whose progenitor grew at both ends (bipolar) always grow at the old end (follow Mitchison's rule).

Nucleoside diphosphokinase, an enzyme with step changes in activity during the cell cycle of the fission yeast Schizosaccharomyces pombe. II. Dissociation of the steps from the DNA-division cycle after induction synchronization

1989 ◽  
Vol 93 (1) ◽  
pp. 185-189
Author(s):  
J. Creanor ◽  
J.M. Mitchison
Keyword(s):  
Cell Cycle ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Doubling Time ◽  
Phase Relations ◽  
Restrictive Temperature ◽  
Permissive Temperature ◽  
Normal Cycle ◽  
Short Cycle ◽  
Cycle Times

Synchrony was induced in cultures of the mitotic mutant cdc2.33 of Schizosaccharomyces pombe by shifting up an asynchronous culture to the restrictive temperature for a period of 3.5-4.5 h and then shifting down to the permissive temperature. The resulting synchronous divisions had short cycle times, down to 50% of the normal cycle. The oscillatory control of nucleoside diphosphokinase activity was also synchronized by the shift-down and the activity rose in a step pattern. Unlike the situation in the normal cycle, this step pattern was dissociated from the shortened cell cycle and had a longer period and different phase relations. It may be that the normal entrainment or coupling between the cell cycle and the activity control fails if the cell cycle is too short. The period of the activity control (equal to the protein doubling time at the restrictive temperature) appears to be temperature-compensated.

scholarly journals The Constitutive Centromere Component CENP-50 Is Required for Recovery from Spindle Damage

2005 ◽  
Vol 25 (23) ◽  
pp. 10315-10328 ◽  
Author(s):  
Yukinori Minoshima ◽  
Tetsuya Hori ◽  
Masahiro Okada ◽  
Hiroshi Kimura ◽  
Tokuko Haraguchi ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Growth Rate ◽  
Mitotic Checkpoint ◽  
Loss Of Function ◽  
Wild Type ◽  
Centromere Function ◽  
Dt40 Cells ◽  
Precise Role ◽  
Chicken Dt40 Cell

ABSTRACT We identified CENP-50 as a novel kinetochore component. We found that CENP-50 is a constitutive component of the centromere that colocalizes with CENP-A and CENP-H throughout the cell cycle in vertebrate cells. To determine the precise role of CENP-50, we examined its role in centromere function by generating a loss-of-function mutant in the chicken DT40 cell line. The CENP-50 knockout was not lethal; however, the growth rate of cells with this mutation was slower than that of wild-type cells. We observed that the time for CENP-50-deficient cells to complete mitosis was longer than that for wild-type cells. Centromeric localization of CENP-50 was abolished in both CENP-H- and CENP-I-deficient cells. Coimmunoprecipitation experiments revealed that CENP-50 interacted with the CENP-H/CENP-I complex in chicken DT40 cells. We also observed severe mitotic defects in CENP-50-deficient cells with apparent premature sister chromatid separation when the mitotic checkpoint was activated, indicating that CENP-50 is required for recovery from spindle damage.

Overexpression Phenotypes of Plk1 and Ndrg2 in Schizosaccharomyces Pombe: Fission Yeast System for Mammalian Gene Study

2005 ◽  
Vol 277-279 ◽  
pp. 1-6 ◽  
Author(s):  
Young Joo Jang ◽  
Young Sook Kil ◽  
Jee Hee Ahn ◽  
Jae Hoon Ji ◽  
Jong Seok Lim ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Rna Splicing ◽  
Mammalian Cells ◽  
Protein Modification ◽  
Genetic System ◽  
Functional Study ◽  
Mammalian Genes

The fission yeast, Schizosaccharomyces pombe is a single-celled free-living fungus that shares many features with cells of more complicated eukaryotes. Many of the genes required for the cell-cycle control, proteolysis, protein modification, and RNA splicing are highly conserved with those of higher eukaryotes. Moreover, fission yeast has the merit of genetics and its genetic system is already well characterized. As such, the current study evaluated the use of a fission yeast system as a tool for the functional study of mammalian genes and attempted to set up an assay system for novel genes. Since the phenotypes of a deletion mutant and the overexpression of a gene are generally analyzed for a functional study of specific genes in yeast, the present study used overexpression phenotypes to study the functions of mammalian genes. Therefore, based on using a thiamine-repressive promoter, two mammalian genes were expressed in fission yeast, and their overexpressed phenotypes compared with those in mammalian cells. The phenotypes resulting from overexpression were analyzed using a FACS, which analyzes the DNA contents, and a microscope. One of the selected genes was the mammalian Polo-like kinase 1 (Plk1), which is activated and plays a role in the mitotic phase of the cell division cycle. The overexpression of various constructs of Plk1 in the HeLa cells caused cell cycle defects, suggesting that the ectopic Plk1s blocked the endogenous Plk1 in the cells. As expected, when the constructs were overexpressed in the fission yeast system, the cells were arrested in mitosis and defected at the end of mitosis. As such, this data suggests that the Plk1-overexpressed phenotypes were similar in the mammalian cells and the fission yeast, thereby enabling the mammalian Plk1 functions to be approximated in the fission yeast. The other selected gene was the N-Myc downstream-regulated gene 2 (ndrg2), which is upregulated during cell differentiation, yet still not well characterized. When the ndrg2 gene was overexpressed in the fission yeast, the cells contained multi-septa. The septa were positioned well, yet their number increased per cell. Therefore, this gene was speculated to block cell division in the last stage of the cell cycle, making the phenotype potentially useful for explaining cell growth and differentiation in mammalian cells. Accordingly, fission yeast is demonstrated to be an appropriate species for the functional study of mammalian genes.

The cell cycle genes cdc22 + and suc22 + of the fission yeast Schizosaccharomyces pombe encode the large and small subunits of ribonucleotide reductase

1993 ◽  
Vol 238-238 (1-2) ◽  
pp. 241-251 ◽  
Author(s):  
Maria-Jose Fernandez Sarabia ◽  
Christopher McInerny ◽  
Pamela Harris ◽  
Colin Gordon ◽  
Peter Fantes
Keyword(s):  
Cell Cycle ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Ribonucleotide Reductase ◽  
Cell Cycle Genes

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.

Polypeptide synthesis in cell cycle mutants of fission yeast

1981 ◽  
Vol 51 (1) ◽  
pp. 203-217
Author(s):  
D.P. Dickinson
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Fission Yeast ◽  
Gel Electrophoresis ◽  
Mutant Cell ◽  
Wild Type ◽  
Unsuccessful Attempt ◽  
Dependent Sequence ◽  
Polypeptide Synthesis ◽  
Cellular Components

The cell cycle of a growing cel is characterized by 3 main periodic events: DNA synthesis mitosis and cell division. These events generally lie in a dependent sequence, in which one event cannot occur unless preceding events have occurred. The existence of dependent sequences of events raises the possibility that at least some of the gene products involved in the events are synthesized in a dependent sequence parallel to the observable events. To test this hypothesis, the patterns of polypeptide synthesis were investigated in 2 types of cell cycle mutant of the fission yeast Schizosaccharomyces pombe: temperature-sensitive cell cycle (ts cdc) mutants. which become blocked in cell cycle progress at the restrictive temperature; and wee I mutants, which are defective in size control over nuclear division, and which divide at a small size. Cells of mutants and wild-type cells were labelled with [35S[sulphate under conditions designed to maximize any differences between the labelling patterns of wild-type and mutant cell polypeptides. The polypeptides were then separated by O'Farrell 2-dimensional gel electrophoresis, and the patterns compared. Although both types of mutation affect cell cycle control, and cause a considerable alteration in the relative proportions of cellular components, an examination of over 700 polypeptides detected on gels revealed no qualitative differences between wild-type and mutant cell polypeptides. These results suggest that a large majority of the more abundant polypeptides in the growing cell are synthesized independently of cell cycle controls directly related to DNA synthesis and division, and that the synthesis of these polypeptides can occur in the absence of normal progress through the cell cycle. Dependent sequences of gene expression do not appear to make a significant contribution to total polypeptide synthesis during the cell cycle, or to the occurrence of periodic cell cycle events such as mitosis. It is suggested that such cell cycle events may result largely through the reorganization of existing cellular components, rather than by the synthesis of new ones. An unsuccessful attempt was made to detect the wee I gene product on gels by surveying a range of mutants for changes in an individual spot. The limitations of gel electrophoresis for this type of survey, and other cell cycle experiments, are discussed.

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