Acetate assimilation by the fission yeast, Schizosaccharomyces pombe

1989 ◽  
Vol 67 (8) ◽  
pp. 464-467 ◽  
Author(s):  
C. S. Tsai ◽  
Ken P. Mitton ◽  
Byron F. Johnson
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Neutral Lipids ◽  
Glyoxylate Cycle ◽  
Yeast Cells ◽  
Peptide Structure ◽  
Synthetase Activity ◽  
Sterol Esters ◽  
Acetate Utilization ◽  
Structure Of Proteins

The fission yeast Schizosaccharomyces pombe utilizes acetate at subinhibitory concentrations in the presence of D-glucose. The nonionized form of acetate is preferentially utilized, oxidized to 14CO2, and assimilated into lipids and proteins. Acetyl CoA synthetase activity greatly increases in the yeast cells grown in media containing acetate. However, glyoxylate cycle enzymes are not detectable in Schizosaccharomyces pombe. [1-14C] Acetate is incorporated into stereols, sterol esters, neutral lipids, and phospholipids. Assimilation of [1-14C]acetate into the peptide structure of proteins was confirmed by a proteolytic digestion experiment.Key words: acetate utilization, fission yeast, Schizosaccharomyces pombe.

Enzyme activities of D-glucose metabolism in the fission yeast Schizosaccharomyces pombe

1992 ◽  
Vol 38 (12) ◽  
pp. 1313-1319 ◽  
Author(s):  
C. S. Tsai ◽  
J.-L. Shi ◽  
B. W. Beehler ◽  
B. Beck
Keyword(s):  
Steady State ◽  
Alcohol Dehydrogenase ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Pyruvate Dehydrogenase ◽  
Pyruvate Dehydrogenase Complex ◽  
Pyruvate Decarboxylase ◽  
Washout Rate ◽  
Synthetase Activity ◽  
Dehydrogenase Complex

The activities of key enzymes that are members of D-glucose metabolic pathways in Schizosaccharomyces pombe undergoing respirative, respirofermentative, and fermentative metabolisms are monitored. The steady-state activities of glycolytic enzymes, except phosphofructokinase, decrease with a reduced efficiency in D-glucose utilization by yeast continuous culture. On the other hand, the enzymic activities of pentose monophosphate pathway reach the maximum when the cell mass production of the cultures is optimum. Enzymes of tricarboxylate cycle exhibit the maximum activities at approximately the washout rate. The steady-state activity of pyruvate dehydrogenase complex increases rapidly when D-glucose is efficiently utilized. By comparison, the activity of pyruvate decarboxylase begins to increase only when ethanol production occurs. Depletion of dissolved oxygen suppresses the activity of pyruvate dehydrogenase complex but facilitates that of pyruvate decarboxylase. Acetate greatly enhances the acetyl CoA synthetase activity. Similarly, ethanol stimulates alcohol dehydrogenase and aldehyde dehydrogenase activities. Evidence for the existence of alcohol dehydrogenase isozymes in the fission yeast is presented. Key words: yeast, glucose-metabolizing enzymes.

scholarly journals Phylogenetic and comparative functional analysis of the cell-separation α-glucanase Agn1p in Schizosaccharomyces

Microbiology ◽  
2014 ◽  
Vol 160 (6) ◽  
pp. 1063-1074 ◽  
Author(s):  
Matthias Sipiczki ◽  
Anita Balazs ◽  
Aniko Monus ◽  
Laszlo Papp ◽  
Anna Horvath ◽  
...  
Keyword(s):  
Cell Wall ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Mother Cell ◽  
Cell Separation ◽  
Yeast Cells ◽  
Glycoside Hydrolase Family ◽  
Binding Domains ◽  
Yeast Phase ◽  
Mother Cell Wall

The post-cytokinetic separation of cells in cell-walled organisms involves enzymic processes that degrade a specific layer of the division septum and the region of the mother cell wall that edges the septum. In the fission yeast Schizosaccharomyces pombe, the 1,3-α-glucanase Agn1p, originally identified as a mutanase-like glycoside hydrolase family 71 (GH71) enzyme, dissolves the mother cell wall around the septum edge. Our search in the genomes of completely sequenced fungi identified GH71 hydrolases in Basidiomycota, Taphrinomycotina and Pezizomycotina, but not in Saccharomycotina. The most likely Agn1p orthologues in Pezizomycotina species are not mutanases having mutanase-binding domains, but experimentally non-characterized hypothetical proteins that have no carbohydrate-binding domains. The analysis of the GH71 domains corroborated the phylogenetic relationships of the Schizosaccharomyces species determined by previous studies, but suggested a closer relationship to the Basidiomycota proteins than to the Ascomycota proteins. In the Schizosaccharomyces genus, the Agn1p proteins are structurally conserved: their GH71 domains are flanked by N-terminal secretion signals and C-terminal sequences containing the conserved block YNFNAY/HTG. The inactivation of the agn1Sj gene in Schizosaccharomyces japonicus, the only true dimorphic member of the genus, caused a severe cell-separation defect in its yeast phase, but had no effect on the hyphal growth and yeast-to-mycelium transition. It did not affect the mycelium-to-yeast transition either, only delaying the separation of the yeast cells arising from the fragmenting hyphae. The heterologous expression of agn1Sj partially rescued the separation defect of the agn1Δ cells of Schizosaccharomyces pombe. The results presented indicate that the fission yeast Agn1p 1,3-α-glucanases of Schizosaccharomyces japonicus and Schizosaccharomyces pombe share conserved functions in the yeast phase.

scholarly journals Intramitotic controls in the fission yeast Schizosaccharomyces pombe: the effect of cell size on spindle length and the timing of mitotic events.

1990 ◽  
Vol 110 (5) ◽  
pp. 1617-1621 ◽  
Author(s):  
I M Hagan ◽  
P N Riddle ◽  
J S Hyams
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Average Length ◽  
Nuclear Migration ◽  
Yeast Cells ◽  
Reverse Direction ◽  
Wild Type ◽  
Spindle Elongation ◽  
Anaphase B ◽  
In Cells

We have used a new cinemicroscopy technique in combination with antitubulin immunofluorescence microscopy to investigate the timing of mitotic events in cells of the fission yeast Schizosaccharomyces pombe having lengths at division between 7 and 60 microns. Wild-type fission yeast cells divide at a length of 14 microns. Separation of daughter nuclei (anaphase B) proceeds at a rate of 1.6 +/- 0.2 microns min-1, until the spindle extends the length of the cell. Coincident with spindle depolymerization, the nuclei reverse direction and take up positions that will become the center of the two daughter cells. This post-mitotic nuclear migration occurs at a rate of 1.4 +/- 0.5 microns-1. In cells in which the weel+ gene is overexpressed fivefold and that have an average length at mitosis of 28 microns, the rate of nuclear separation was only slightly reduced but, as spindles in these cells measure 20-22 microns, the duration of anaphase B was extended by approximately 40%. By contrast, in the mutant weel.50, which divides at 7 microns, both the rate and duration of anaphase B were indistinguishable from wild type. Nuclei reach the ends of these cells earlier but remain there until a point corresponding to the time of postmitotic nuclear migration in wild type. Thus, the events of mitosis can be extended but not abbreviated. These results are discussed in terms of a mitotic termination control that monitors many different events, one of which is spindle elongation.

scholarly journals Histone H3 K36 Methylation Is Associated with Transcription Elongation in Schizosaccharomyces pombe

2005 ◽  
Vol 4 (8) ◽  
pp. 1446-1454 ◽  
Author(s):  
Stephanie A. Morris ◽  
Yoichiro Shibata ◽  
Ken-ichi Noma ◽  
Yuko Tsukamoto ◽  
Erin Warren ◽  
...  
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Rna Polymerase Ii ◽  
Budding Yeast ◽  
Transcription Elongation ◽  
Synthetic Medium ◽  
Histone H3 ◽  
Yeast Cells ◽  
Pol Ii ◽  
Elongation Process

ABSTRACT Set2 methylation of histone H3 at lysine 36 (K36) has recently been shown to be associated with RNA polymerase II (Pol II) elongation in Saccharomyces cerevisiae. However, whether this modification is conserved and associated with transcription elongation in other organisms is not known. Here we report the identification and characterization of the Set2 ortholog responsible for K36 methylation in the fission yeast Schizosaccharomyces pombe. We find that similar to the budding yeast enzyme, S. pombe Set2 is also a robust nucleosome-selective H3 methyltransferase that is specific for K36. Deletion of the S. pombe set2 + gene results in complete abolishment of K36 methylation as well as a slow-growth phenotype on plates containing synthetic medium. These results indicate that Set2 is the sole enzyme responsible for this modification in fission yeast and is important for cell growth under stressed conditions. Using the chromatin immunoprecipitation assay, we demonstrate that K36 methylation in S. pombe is associated with the transcribed regions of Pol II-regulated genes and is devoid in regions that are not transcribed by Pol II. Consistent with a role for Set2 in transcription elongation, we find that S. pombe Set2 associates with the hyperphosphorylated form of Pol II and can fully rescue K36 methylation and Pol II interaction in budding yeast cells deleted for Set2. These results, along with our finding that K36 methylation is highly conserved among eukaryotes, imply a conserved role for this modification in the transcription elongation process.

scholarly journals Schizosaccharomyces pombe Noc3 Is Essential for Ribosome Biogenesis and Cell Division but Not DNA Replication

2008 ◽  
Vol 7 (9) ◽  
pp. 1433-1440 ◽  
Author(s):  
Christopher R. Houchens ◽  
Audrey Perreault ◽  
François Bachand ◽  
Thomas J. Kelly
Keyword(s):  
Cell Division ◽  
Dna Replication ◽  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Ribosome Biogenesis ◽  
Budding Yeast ◽  
Ribosomal Subunit ◽  
Dna Helicase ◽  
Yeast Cells ◽  
Direct Role

ABSTRACT The initiation of eukaryotic DNA replication is preceded by the assembly of prereplication complexes (pre-RCs) at chromosomal origins of DNA replication. Pre-RC assembly requires the essential DNA replication proteins ORC, Cdc6, and Cdt1 to load the MCM DNA helicase onto chromatin. Saccharomyces cerevisiae Noc3 (ScNoc3), an evolutionarily conserved protein originally implicated in 60S ribosomal subunit trafficking, has been proposed to be an essential regulator of DNA replication that plays a direct role during pre-RC formation in budding yeast. We have cloned Schizosaccharomyces pombe noc3 + (Spnoc3 +), the S. pombe homolog of the budding yeast ScNOC3 gene, and functionally characterized the requirement for the SpNoc3 protein during ribosome biogenesis, cell cycle progression, and DNA replication in fission yeast. We showed that fission yeast SpNoc3 is a functional homolog of budding yeast ScNoc3 that is essential for cell viability and ribosome biogenesis. We also showed that SpNoc3 is required for the normal completion of cell division in fission yeast. However, in contrast to the proposal that ScNoc3 plays an essential role during DNA replication in budding yeast, we demonstrated that fission yeast cells do enter and complete S phase in the absence of SpNoc3, suggesting that SpNoc3 is not essential for DNA replication in fission yeast.

scholarly journals Lipid Composition of Sheffersomyces stipitis M12 Strain Grown on Glycerol as a Carbon Source

2020 ◽  
Vol 58 (2) ◽  
pp. 203-213
Author(s):  
Stela Križanović ◽  
Damir Stanzer ◽  
Gordana Čanadi Jurešić ◽  
Elizabeta Kralj ◽  
Karla Hanousek-Čiča ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Lipid Metabolism ◽  
Carbon Source ◽  
Lipid Composition ◽  
Solid Phase ◽  
Neutral Lipids ◽  
Wild Strain ◽  
Yeast Cells ◽  
Sterol Esters ◽  
Ergosterol Synthesis

Research background. In this study the content and composition of lipids in ergosterol-reduced Sheffersomyces stipitis M12 strain grown on glycerol as a carbon source is determined. Blocking the ergosterol synthesis route in yeast cells is a recently proposed method for increasing S-adenosyl-L-methionine (SAM) production. Experimental approach. The batch cultivation of M12 yeast was carried out under aerobic conditions in a laboratory bioreactor with glycerol as carbon source, and with pulsed addition of methionine. Glycerol and SAM content were monitored by high-performance liquid chromatography, while fatty acid composition of different lipid classes, separated by solid phase extraction, was determined by gas chromatography. Results and conclusion. Despite the reduced amount of ergosterol in yeast cells, thanks to the reorganized lipid metabolism, M12 strain achieved high biomass yield and SAM production. Neutral lipids prevailed (making more than 75 % of total lipids), but their content and composition differed significantly in the two tested types of yeast. Unsaturated and C18 fatty acids prevailed in both the M12 strain and wild type. In all fractions except free fatty acids, the index of unsaturation in M12 strain was lower than in the wild strain. Our tested strain adjusts itself by changing the content of lipids (mainly phospholipids, sterols and sterol esters), and with desaturation adjustments, to maintain proper functioning and fulfil increased energy needs.Novelty and scientific contribution. Reorganization of S. stipitis lipid composition caused by blocking the metabolic pathway of ergosterol synthesis was presented. A simple scheme of actual lipid metabolism during active SAM production in S. stipitis, grown on glycerol was constructed and shown. This fundamental knowledge of lipid metabolic pathways will be a helpful tool in improving S. stipitis as an expression host and a model organism, opening new perspectives for its applied research.

Pseudo-exponential growth in length of the fission yeast, Schizosaccharomyces pombe

1988 ◽  
Vol 34 (12) ◽  
pp. 1338-1343 ◽  
Author(s):  
Hisao Miyata ◽  
Machiko Miyata ◽  
Byron F. Johnson
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Exponential Growth ◽  
Linear Growth ◽  
Normal Growth ◽  
Growth Patterns ◽  
Birth Length ◽  
Yeast Cells ◽  
Wild Type ◽  
Constant Length

The growth patterns of individual cells of the fission yeast (Schizosaccharomyces pombe wild-type cells, strain 972 h−; cells exposed to hydroxyurea; and cdc mutants, 11-123, 2-33) were investigated by time-lapse photomicrography. Wild-type cells showed one, two, or three linear-growth segments followed by a constant-length stage. Cells with two segments were most frequent. Hydroxyurea cells that divided as oversized cells (about three times the birth length) had three linear-growth segments in a cycle. Mutant cdc11-123 cells did not divide but had a constant-length stage separating the cycles; both the first and second cycles consisted of two linear-growth segments, and cells were oversized at the second constant-length stage (about 3.5 times the birth length). Elongating cdc2-33 cells that did not divide and were oversized (about five times the birth length) while under observation, showed four linear-growth segments. Cells of all strains showed 30 to 40% increase in growth rate at the rate-change point and maintained approximate exponential (pseudo-exponential) growth. We conclude that the normal growth pattern of individual fission-yeast cells is the pseudo-exponential pattern.

scholarly journals Degradation of HMG-CoA reductase-induced membranes in the fission yeast, Schizosaccharomyces pombe.

1995 ◽  
Vol 131 (1) ◽  
pp. 81-94 ◽  
Author(s):  
P Y Lum ◽  
R Wright
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Neutral Lipids ◽  
Degradation Process ◽  
Biosynthetic Enzyme ◽  
Common Pathway ◽  
Hmg Coa Reductase ◽  
Lipid Particle ◽  
Coa Reductase ◽  
New Protein

Elevated levels of certain membrane proteins, including the sterol biosynthetic enzyme HMG-CoA reductase, induce proliferation of the endoplasmic reticulum. When the amounts of these proteins return to basal levels, the proliferated membranes are degraded, but the molecular details of this degradation remain unknown. We have examined the degradation of HMG-CoA reductase-induced membranes in the fission yeast, Schizosaccharomyces pombe. In this yeast, increased levels of the Saccharomyces cerevisiae HMG-CoA reductase isozyme encoded by HMG1 induced several types of membranes, including karmellae, which formed a cap of stacked membranes that partially surrounded the nucleus. When expression of HMG1 was repressed, the karmellae detached from the nucleus and formed concentric, multilayered membrane whorls that were then degraded. During the degradation process, CDCFDA-stained compartments distinct from preexisting vacuoles formed within the interior of the whorls. In addition to these compartments, particles that contained neutral lipids also formed within the whorl. As the thickness of the whorl decreased, the lipid particle became larger. When degradation was complete, only the lipid particle remained. Cycloheximide treatment did not prevent the formation of whorls. Thus, new protein synthesis was not needed for the initial stages of karmellae degradation. On the contrary, cycloheximide promoted the detachment of karmellae to form whorls, suggesting that a short lived protein may be involved in maintaining karmellae integrity. Taken together, these results demonstrate that karmellae membranes differentiated into self-degradative organelles. This process may be a common pathway by which ER membranes are turned over in cells.

scholarly journals Increase in cellular triacylglycerol content and emergence of large ER-associated lipid droplets in the absence of CDP-DG synthase function

2014 ◽  
Vol 25 (25) ◽  
pp. 4083-4095 ◽  
Author(s):  
Yue He ◽  
Candice Yam ◽  
Kyle Pomraning ◽  
Jacqueline S. R. Chin ◽  
Joanne Y. Yew ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Phosphatidic Acid ◽  
Fission Yeast ◽  
Lipid Droplets ◽  
Neutral Lipids ◽  
Sterol Esters ◽  
Triacylglycerol Synthesis ◽  
Kennedy Pathway ◽  
Triacylglycerol Content ◽  
Cellular Organelles

Excess fatty acids and sterols are stored as triacylglycerols and sterol esters in specialized cellular organelles, called lipid droplets. Understanding what determines the cellular amount of neutral lipids and their packaging into lipid droplets is of fundamental and applied interest. Using two species of fission yeast, we show that cycling cells deficient in the function of the ER-resident CDP-DG synthase Cds1 exhibit markedly increased triacylglycerol content and assemble large lipid droplets closely associated with the ER membranes. We demonstrate that these unusual structures recruit the triacylglycerol synthesis machinery and grow by expansion rather than by fusion. Our results suggest that interfering with the CDP-DG route of phosphatidic acid utilization rewires cellular metabolism to adopt a triacylglycerol-rich lifestyle reliant on the Kennedy pathway.

Sexual co-flocculation and asexual self-flocculation of heterothallic fission-yeast cells(Schizosaccharomyces pombe)

1987 ◽  
Vol 33 (8) ◽  
pp. 684-688 ◽  
Author(s):  
Byron F. Johnson ◽  
Teena Walker ◽  
Machiko Miyata ◽  
Hisao Miyata ◽  
G. B. Calleja
Keyword(s):  
Schizosaccharomyces Pombe ◽  
Fission Yeast ◽  
Yeast Cells ◽  
Logarithmic Phase ◽  
N Sources ◽  
Late Logarithmic Phase ◽  
Culturing Conditions

Culturing conditions have been found that engender flocculation of strains 975h+ and 972h− of the fission yeast Schizosaccharomyces pombe Lindner. Both strains separately exhibit asexual self-flocculation and together, sexual co-flocculation. Flocculation is glucose repressible, but that repression is leaky, because self-flocculation can occur during the late logarithmic phase of growth before either of the C or N sources are depleted.

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