Action of Ribonuclease Preparations on Viable Yeast Cells and Spheroplasts

Schlenk, F.; Dainko, J. L.

doi:10.1128/jb.89.2.428-436.1965

Cited by 44 publications

(29 citation statements)

References 21 publications

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“…This was confirmed by the observation that the poly-L-lysine effect was not inhibited by sodium chloride concentrations up to 25 mM. As shown by Schlenk and Dainko (1965) and Yphantis et al…”

Section: Discussionsupporting

confidence: 58%

An assay of relative cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe

Nobel

Klis

Munnik

et al. 1990

Yeast

144

113

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We have developed a new assay to determine relative cell wall porosity in yeasts, which is based on polycation-induced leakage of UV-absorbing compounds. Polycations with a small hydrodynamic radius as measured by gel filtration (poly-L-lysine) caused cell leakage independent of cell wall porosity whereas polycations with a large hydrodynamic radius (DEAE-dextrans) caused only limited cell leakage due to limited passage through the cell wall. This allowed the ratio between DEAE-dextran- and poly-L-lysine-induced cell leakage to be used as a measure of cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe. Using this assay, we found that the composition of the growth medium affected cell wall porosity in S. cerevisiae. In addition, we could show that cell wall porosity is limited by the number of disulphide bridges in the wall and is dependent on cell turgor. It is argued that earlier methods to estimate cell wall porosity in S. cerevisiae resulted in large underestimations.

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Section: Discussionsupporting

confidence: 58%

An assay of relative cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe

Nobel

Klis

Munnik

et al. 1990

Yeast

144

113

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“…Growth of S. cerevisiae at 15 C instead of 30 C also leads to synthesis of an increased proportion of phosphatidylcholine (5,12), and our data indicate that the greater cation-binding power which this may confer on the membrane (5) also does not affect retention of membrane proteins when the cells are subjected to cold osmotic shock. The ability of S. cerevisiae cells to lose membrane proteins as a result of cold osmotic shock may be related to the phenomena reported by Schlenk and his colleagues (32,33); they found that Candida utilis and S. cerevisiae, in the absence of salts and buffers, become sensitive to several proteins of moderate size, including ribonuclease, protamine, lysozyme, and bovine plasma albumin. The sensitivity of these cells to proteins could be explained by the ability of the proteins to replace membrane-bound proteins in cation-free suspensions and might explain the need for EDTA ii the cold osmotic shock-induced loss of glucosamine-accumulating ability.…”

Section: Discussionmentioning

confidence: 75%

Cold Osmotic Shock in Saccharomyces cerevisiae

Patching

Rose

1971

J Bacteriol

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Saccharomyces cerevisiae NCYC 366 is susceptible to cold osmotic shock. Exponentially growing cells from batch cultures grown in defined medium at 30 C, after being suspended in 0.8 M mannitol containing 10 mm ethylenediaminetetraacetic acid and then resuspended in ice-cold 0.5 mM MgCl2, accumulated the nonmetabolizable solutes D-glucosamine-hydrochloride and 2-aminoisobutyrate at slower rates than unshocked cells; shocked cells retained their viability. Storage of unshocked batch-grown cells in buffer at 10 C led to an increase in ability to accumulate glucosamine, and further experiments were confined to cells grown in a chemostat under conditions of glucose limitation, thereby obviating the need for storing cells before use. A study was made of the effect of the different stages in the cold osmotic shock procedure, including the osmotic stress, the chelating agent, and the cold Mg2+-containing diluent, on viability and solute-accumulating ability. Growth of shocked cells in defined medium resembled that of unshocked cells; however, in malt extract-yeast extract-glucose-peptone medium, the shocked cells had a longer lag phase of growth and initially grew at a slower rate. Cold osmotic shock caused the release of low-molecular-weight compounds and about 6 to 8% of the cell protein. Neither the cell envelope enzymes, invertase, acid phosphatase and L-leucine-f,-naphthylamidase, nor the cytoplasmic enzyme, alkaline phosphatase, were released when yeast cells were subjected to cold osmotic shock.A number of different shock treatments cause the release from microbial cells of low-and highmolecular-weight compounds, effects which are often accompanied by death of the cells. One of the first shock treatments to be studied in detail was referred to as "cold shock," in which sudden chilling of a dilute suspension of exponential-phase gram-negative bacteria in water or very dilute buffer causes the death of many of the cells (6,9,10,20,34,36,37). Cold shock, which is manifested mainly in gram-negative (36, 37) bacteria, is accompanied by release of intracellular amino acids and nucleotides (36). Cold shock has not been reported in yeasts (20). The main modification to the cold shock treatment has consisted in incubating cells in a hyperosmolar solution of inert solute (e.g. sucrose) containing ethylenediaminetetraacetic acid (EDTA) followed by centrifugation and suspending the cells in cold water containing Mg2+. This treatment has been termed osmotic shock; the earlier literature on it was reviewed by Heppel ( 11). Osmotic shock, which strictly speaking should be referred to as cold osmotic shock (40), resembles cold shock in that it causes release of low-and high-molecular-weight compounds, including periplasmic enzymes, such as acid phosphatase (27) and nucleotidases (23, 24), and certain mem-brane-bound transport proteins (13, 28); however, it does not usually lead to a decrease in the viability of the cell population. Cold osmotic shock has been demonstrated in gram-negative bacteria, especially in members of the En...

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“…Samples of incubated cells (2 ml) were filtered on a 0.22-pm Millipore filter and the absorbance of filtrates was measured at 260 and 280 nm. The maximum extractable UV absorbing materials was determined after treatment of cells with 2.2 M perchloric acid at 100°C for 30 min; this value was taken as 100% release [12].…”

Section: Leakage Of Uv Absorbing Materialsmentioning

confidence: 99%

Effect of the lipopeptide antibiotic, iturin A, on morphology and membrane ultrastructure of yeast cells

Thimon¹,

Peypoux²,

Wallach³

et al. 1995

FEMS Microbiology Letters

115

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The effects of iturin A, at fungicidal concentrations, on yeast cells were studied by scanning electron microscopy and by transmission electron microscopy. A depression, observed in each iturin A-treated cell, was the consequence of the release of electrolytes and other cytoplasmic components. Iturin A passes through the cell wall and disrupts the plasma membrane with the formation of small vesicles and the aggregation of intramembranous particles. Moreover, iturin A passes through the plasma membrane and interacts with the nuclear membrane and probably with membranes of other cytoplasmic organelles.

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Action of Ribonuclease Preparations on Viable Yeast Cells and Spheroplasts

Cited by 44 publications

References 21 publications

An assay of relative cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe

An assay of relative cell wall porosity in Saccharomyces cerevisiae, Kluyveromyces lactis and Schizosaccharomyces pombe

Cold Osmotic Shock in Saccharomyces cerevisiae

Effect of the lipopeptide antibiotic, iturin A, on morphology and membrane ultrastructure of yeast cells

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