Metabolic state and cell cycle as determinants of facilitated uptake of genetic information by yeast Saccharomyces cerevisiae

Chaustova, Larisa; Miliukienė, Valė; Zimkus, Aurelijus; Razumas, Valdemaras

doi:10.2478/s11535-008-0040-7

Cited by 6 publications

(4 citation statements)

References 19 publications

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“…Figure 2 shows that the steady state was not reached even after incubation for 100 min. Our results are in agreement with other observations where the steady state was reached after 15-100 min [Ballarin-Denti et al, 1994;Chaustova et al, 2008]. These results indicate that the negatively charged surfaces of the yeast cell walls [Jigami and Odani, 1999;Klis et al, 2006] may play an important role in preventing the accumulation of cationic compounds such as TPP þ into the cell.…”

Section: Discussionsupporting

confidence: 93%

Electric field‐induced effects on yeast cell wall permeabilization

Stirkė

Zimkus

Ramanavičienė

et al. 2013

Bioelectromagnetics

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The permeability of the yeast cells (Saccharomyces cerevisiae) to lipophilic tetraphenylphosphonium cations (TPP(+) ) after their treatment with single square-shaped strong electric field pulses was analyzed. Pulsed electric fields (PEF) with durations from 5 to 150 µs and strengths from 0 to 10 kV/cm were applied to a standard electroporation cuvette filled with the appropriate buffer. The TPP(+) absorption process was analyzed using an ion selective microelectrode (ISE) and the plasma membrane permeability was determined by measurements obtained using a calcein blue dye release assay. The viability of the yeast and the inactivation of the cells were determined using the optical absorbance method. The experimental data taken after yeasts were treated with PEF and incubated for 3 min showed an increased uptake of TPP(+) by the yeast. This process can be controlled by setting the amplitude and pulse duration of the applied PEF. The kinetics of the TPP(+) absorption process is described using the second order absolute rate equation. It was concluded that the changes of the charge on the yeast cell wall, which is the main barrier for TPP(+) , is due to the poration of the plasma membrane. The applicability of the TPP(+) absorption measurements for the analysis of yeast cells electroporation process is also discussed.

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Section: Discussionsupporting

confidence: 93%

Electric field‐induced effects on yeast cell wall permeabilization

Stirkė

Zimkus

Ramanavičienė

et al. 2013

Bioelectromagnetics

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“…One interesting report on PEG-dependent transformation of otherwise nonmanipulated intact yeast cells was also foreseen as a natural transformation strongly correlated with the growth phase [25]. Such competence can be induced/enhanced mechanically since it may involve concomitant cell wall damage due to heat shock [12,25,76] or pH shift [25]. Both mechanical damage and heat shock seem relevant environmental conditions, and pH shift is already known to be important in yeast transfection [54].…”

Section: Environmentally Induced Yeast Competencementioning

confidence: 99%

“…1) and might be open during mitosis when nucleus is being remodeled and thus possess more loosen membrane structure [32] supporting entrance of the endosomically released tDNA or, alternatively, endosome-nuclear membrane fusion which delivers the engulfed tDNA directly [38]. Indeed, the highest artificial transformability was seen at S phase [12] when envelope changes enable continuous nuclear expansion [32]. Cells with prolonged S phase were proven most competent contrary to cells arrested at M-phase [11] (no data for G 1 arrested cells).…”

Section: Mechanisms Routes and Pathways Of Edecmentioning

confidence: 99%

Ecologically Driven Competence for Exogenous DNA Uptake in Yeast

Mitrikeski

2015

Curr Microbiol

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Unlike prokaryotes, eukaryotic organisms do not seem to be equipped with natural cell process(es) designated for exogenous DNA uptake. However, it is barely known that under laboratory circumstances resembling wild fungal environment(s), at least some lower eukaryotes could become naturally competent for exogenous DNA uptake. Thus, apart from the known fact that non-manipulated cells of yeast Saccharomyces cerevisiae take exogenous DNA by conjugation with certain bacteria, there are also mechanical and physiological mechanisms enabling their transformation under environmental conditions. This clearly shows that lower eukaryotes are amenable to transformation without applying man-made technology (i.e., naturally). However, this topic failed to raise critical scientific interest. Therefore, this review aims to scrutinize the overall implication of the phenomenon stressing its fundamental and applicable importance. It also summarizes all axiomatic laboratory circumstances/vehicles hitherto known to provoke yeast competence naturally and critically discusses plausible mechanisms behind. Possible pathways underlying the phenomenon are emphasized and a unifying model is proposed. This story potentially spans several different research fields, from evolutionary genetics to genetic transformation technology.

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“…Li + ions enhance the transformation of intact cells, but no effect is observed on transformation of protoplasts, implying that Li + ions facilitate DNA passage through the cell wall [5]. The mechanism underlying S. cerevisiae transformation includes DNA attachment and penetration through the cell wall, although how DNA passes through the cell wall is not yet clear [6][7][8]. Cell wall density, thickness and structure are factors of major importance during penetration of exogenous molecules into the cell.…”

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confidence: 99%

Li+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FT-IR spectroscopy

2013

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+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FT-IR spectroscopy IntroductionThe capacity of metal cations to induce competence of yeast and bacterial cells to exogenous DNA is the basis of biological methods in biotechnology and molecular genetics [1][2][3]. The most commonly used, efficient protocol, for genetic transformation of Saccharomyces cerevisiae is treatment with alkali metal ions. Li + ions are the most effective of all the cations tested, and the transformation efficiency is comparable to levels obtained by the protoplast method [4]. Li + ions enhance the transformation of intact cells, but no effect is observed on transformation of protoplasts, implying that Li + ions facilitate DNA passage through the cell wall [5]. The mechanism underlying S. cerevisiae transformation includes DNA attachment and penetration through the cell wall, although how DNA passes through the cell wall is not yet clear [6][7][8]. Cell wall density, thickness and structure are factors of major importance during penetration of exogenous molecules into the cell. However, despite the extensive use of Li + ions in yeast transformation protocols, the influence of these cations on the structure of the yeast cell wall has not been widely investigated and, therefore, remains unclear. The influence of Li + ions on the cell surface topography of intact S. cerevisiae cells was observed by atomic force microscopy and it was found that the surface of Li + treated yeast cells became much rougher [9].The S. cerevisiae cell wall is composed of three major components: β-glucans, chitin, and mannoproteins. Glucose residues are linked to other glucose molecules through β(1→3) and β(1→6) linkages and to N-acetylglucosamine via β(1→4) bonds [10,11]. S. cerevisiae mannan has a linear α(1→6)-linked Keywords: FT-IR spectroscopy • Lithium • Saccharomyces cerevisiae • TransformationAbstract: The effect of Li + ions as a transformation inducing agent on the yeast cell wall has been studied. Two Saccharomyces cerevisiae strains, p63-DC5 with a native cell wall, and strain XCY42-30D(mnn1) which contains structural changes in the mannan-protein complex, were used. Fourier transform infrared (FT-IR) spectroscopy has been used for the characterization of the yeast strains and for determination of the effect of lithium cations on the cell wall. A comparison of the carbohydrate absorption band positions in the 970-1185 cm -1 range, of Na + and Li + treated yeast cells has been estimated. Absorption band positions of the cell wall carbohydrates of p63-DC5 were not influenced by the studied ions. On the contrary, the treatment of XCY42-30D(mnn1) cells with Li + ions shifted glucan band positions, implying that the cell wall structure of strain XCY42-30D(mnn1) is more sensitive to Li + ion treatment. backbone with side chains of α(1→2)-and α(1→3)-linked mannose units. In S. cerevisiae, mannoproteins contribute to the regulation of cell wall porosity, and therefore control both the exit of secretary proteins and the entrance of ma...

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Metabolic state and cell cycle as determinants of facilitated uptake of genetic information by yeast Saccharomyces cerevisiae

Cited by 6 publications

References 19 publications

Electric field‐induced effects on yeast cell wall permeabilization

Electric field‐induced effects on yeast cell wall permeabilization

Ecologically Driven Competence for Exogenous DNA Uptake in Yeast

Li+ effect on the cell wall of the yeast Saccharomyces cerevisiae as probed by FT-IR spectroscopy

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