A Chromatin Fiber Analysis Pipeline to Model DNA Synthesis and Structures in Fission Yeast

2017 ◽  
pp. 509-526
Author(s):  
Sarah A. Sabatinos ◽  
Marc D. Green
Keyword(s):  
Dna Synthesis ◽  
Fission Yeast ◽  
Chromatin Fiber ◽  
Analysis Pipeline ◽  
Fiber Analysis

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.

Oxygen uptake during the cell cycle of the fission yeast Schizosaccharomyces pombe

1978 ◽  
Vol 33 (1) ◽  
pp. 399-411
Author(s):  
J. Creanor
Keyword(s):  
Cell Cycle ◽  
Cell Division ◽  
Oxygen Uptake ◽  
Dna Synthesis ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Nuclear Division ◽  
Synchronous Culture ◽  
Synchronous Cultures ◽  
The Many

Oxygen uptake was measured in synchronous cultures of the fission yeast Schizosaccharomyces pombe. The rate of oxygen uptake was found to increase in a step-wise manner at the beginning of the cycle and again in the middle of the cycle. The increases in rate were such that overall, oxygen uptake doubled in rate once per cell cycle. Addition of inhibitors of DNA synthesis or nuclear division to a synchronous culture did not affect the uptake of oxygen. In an induced synchronous culture, in which DNA synthesis, cell division, and nuclear division, but not ‘growth’ were synchronized, oxygen uptake increased continuously in rate and did not show the step-wise rises which were shown in the selection-synchronized culture. These results were compared with previous measurements of oxygen uptake in yeast and an explanation is suggested for the many different patterns which have been reported.

Human p53 Restores DNA Synthesis Control in Fission Yeast

1997 ◽  
Vol 378 (11) ◽  
Author(s):  
Matthias Bureik ◽  
Andreas Jungbluth ◽  
Rudolf Drescher ◽  
Peter Wagner
Keyword(s):  
Dna Synthesis ◽  
Fission Yeast

scholarly journals Cyclin-Dependent Kinase Inhibits Reinitiation of a Normal S-Phase Program during G2 in Fission Yeast

2009 ◽  
Vol 29 (15) ◽  
pp. 4025-4032 ◽  
Author(s):  
Lee Kiang ◽  
Christian Heichinger ◽  
Stephen Watt ◽  
Jürg Bähler ◽  
Paul Nurse
Keyword(s):  
Cell Cycle ◽  
Dna Synthesis ◽  
Fission Yeast ◽  
Cell Size ◽  
S Phase ◽  
Cyclin Dependent Kinase

ABSTRACT To achieve faithful replication of the genome once in each cell cycle, reinitiation of S phase is prevented in G2 and origins are restricted from refiring within S phase. We have investigated the block to rereplication during G2 in fission yeast. The DNA synthesis that occurs when G2/M cyclin-dependent kinase (CDK) activity is depleted has been assumed to be repeated rounds of S phase without mitosis, but this has not been demonstrated to be the case. We show here that on G2/M CDK depletion in G2, repeated S phases are induced, which are correlated with normal G1/S transcription and attainment of doublings in cell size. Mostly normal mitotic S-phase origins are utilized, although at different efficiencies, and replication is essentially equal across the genome. We conclude that CDK inhibits reinitiation of S phase during G2, and if G2/M CDK is depleted, replication results from induction of a largely normal S-phase program with only small differences in origin usage and efficiency.

scholarly journals A zinc finger protein controls the onset of premeiotic DNA synthesis of fission yeast in a Mei2-independent cascade.

1994 ◽  
Vol 13 (8) ◽  
pp. 1881-1887 ◽  
Author(s):  
A. Sugiyama ◽  
K. Tanaka ◽  
K. Okazaki ◽  
H. Nojima ◽  
H. Okayama
Keyword(s):  
Dna Synthesis ◽  
Fission Yeast ◽  
Zinc Finger ◽  
Zinc Finger Protein ◽  
Finger Protein ◽  
Independent Cascade

Arginase and sucrase potential in the fission yeast Schizosaccharomyces pombe

1980 ◽  
Vol 46 (1) ◽  
pp. 399-431
Author(s):  
T. Benitez ◽  
P. Nurse ◽  
J.M. Mitchison
Keyword(s):  
Dna Replication ◽  
Dna Synthesis ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Gene Dosage ◽  
Critical Ratio ◽  
Restrictive Temperature ◽  
Wild Type ◽  
Synchronous Cultures ◽  
Short Period

The induction potentials of 2 enzymes, sucrase and arginase, have been measured in asynchronous and synchronous cultures of the fission yeast Schizosaccharomyces pombe. The effect on potential of inhibiting DNA synthesis is asynchronous cultures has been studied using 2 temperature-sensitive dcd mutants, one blocked in DNA replication and the other blocked in mitosis. The results show that despite inhibition of DNA synthesis, sucrase and arginase potential both continue to increase exponentially for at least a generation of growth after shifting the cdc mutants from the permissive to the restrictive temperature. A second method of inhibiting DNA synthesis, using deoxyadenosine, has also been tested. Cells treated with deoxyadenosine stop the increase in potential for a short period. However, experiments carried out using a cdc mutant together with deoxyadenosine show that the block to the increase in potential is due to a side effect of the inhibitor. It appears that increase in potential is not dependent upon continued DNA replication, and that gene dosage does not control potential in the normal cell cycle. This conclusion is supported by measurements on mutants of different cell sizes. potential is proportional to size (protein content per cell is asynchronous culture) and not to DNA content. Although potential is not gene limited in normal cells, it does appear to be so in the abnormally large cells produced by a cdc block. If cdc mutants of different sizes are grown asynchronously, and DNA synthesis is inhibited by a shift to the restrictive temperature, there is no increase in potential. This critical ratio is different for the 2 enzymes, but for each enzyme it is similar in all the mutants tested. When large cells (produced by a mutant block for 4.5 h) are shifted down in temperature, there are synchronous rounds of DNA synthesis and division and also step doublings in potential. In synchronous cultures of wild type cells, both enzymes show a stepwise doubling of potential at 0.2 of a cycle after DNA replication. In synchronous cultures of cdc mutants blocked either in replication or in mitosis, the potential steps continue with the normal timing observed in wild type cells. This shows that the steps are not dependent on the events of the DNA-division cycle but are controlled by another mechanism. Attainment of a critical size might be part of this mechanism, but tests with size mutants argue against this.

Sign in / Sign up

Export Citation Format

Share Document