Ethidium bromide induced mutation of yeast mitochondria: Complete transformation of cells into respiratory deficient non-chromosomal “petites”

Słonimski, Piotr P.; Perrodin, G.; Croft, J. H.

doi:10.1016/0006-291x(68)90440-3

Cited by 489 publications

(136 citation statements)

References 5 publications

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“…RFLP analysis of their mtDNA confirmed the mitochondria1 origin of this respiratory deficiency. Six of the mutants exhibited a complete loss of the mtDNA, analogous to rho' 'petite' mutants of S. cerevisiae, whereas four other mutants showed partial deletions of the mtDNA comparable to 'petite' rho-of S. cerevisiae [34]. In addition, the induction of the complete loss of mtDNA by ethidium bromide treatment of a respiratory competent strain suggested that this resistance was a consequence of the respiratory deficiency.…”

Section: Discussionmentioning

confidence: 93%

In-vitro resistance to azoles associated with mitochondrial DNA deficiency in Candida glabrata

Defontaine

Bouchara

Declerk

et al. 1999

Journal of Medical Microbiology

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A commercially available disk diffusion procedure was used in a large-scale study to evaluate the susceptibility of a wide range of Candida isolates to polyenes and azoles. With almost all isolates of C. glabrata resistant colonies were present within the inhibition zones for the azole compounds fluconazole, ketoconazole and miconazole, and less frequently for isoconazole, econazole and clotrimazole. Ten randomly selected isolates were cloned by limiting dilution and the susceptibility of the resulting strains to polyenes and azoles was determined. All strains presented a similar susceptibility pattern with sensitivity to polyenes and the presence of resistant colonies for all azole compounds except tioconazole. For each strain and each antifungal agent, one of these resistant colonies was subcultured and studied for antifungal susceptibility. All these colonies showed similar properties regardless of which antifungal agent allowed their selection, with increased sensitivity to polyenes and cross-resistance to the azole compounds except tioconazole. Similar results were obtained on Shadomy's modified medium and on synthetic medium. Likewise, determination of MICs by the Etest method confirmed the resistance to fluconazole. Comparative growth studies revealed a respiratory deficiency in the mutants caused by mitochondria1 DNA (mtDNA) deletions. In addition, 'petite' mutants were obtained from a wild-type strain by exposure to ethidium bromide, and these respiratory mutants were shown to be resistant to azoles. These results demonstrate the relationship between mtDNA deficiency and resistance to azoles, and provide an interesting model to study the mechanisms of action of these antifungal agents.

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Section: Discussionmentioning

confidence: 93%

In-vitro resistance to azoles associated with mitochondrial DNA deficiency in Candida glabrata

Defontaine

Bouchara

Declerk

et al. 1999

Journal of Medical Microbiology

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“…After a 60-min chase, greater than 70% of the original Cox2p remained in these strains. In (Slonimski et al, 1968 reduced to approximately 8% of the original level after a 60-min chase ( Figure 5B). Cox2p was stabilized in the cox4 ymel double mutant, such that the level of protein was greater than 90% of the original level after a 60-min chase ( Figure 5B).…”

Section: Orientation Of Ymelp In the Mitochondrialmentioning

confidence: 98%

Biochemical and functional analysis of the YME1 gene product, an ATP and zinc-dependent mitochondrial protease from S. cerevisiae.

Weber

Hanekamp

Thorsness

1996

MBoC

129

106

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Inactivation of YME1 in yeast causes several distinct phenotypes: an increased rate of DNA escape from mitochondria, temperature-sensitive growth on nonfermentable carbon sources, extremely slow growth when mitochondrial DNA is completely absent from the cell, and altered morphology of the mitochondrial compartment. The protein encoded by YME1, Ymelp, contains two highly conserved sequence elements, one implicated in the binding and hydrolysis of ATP, and the second characteristic of active site residues found in neutral, zinc-dependent proteases. Both the putative ATPase and zinc-dependent protease elements are necessary for the function of Ymelp as genes having mutations in critical residues of either of these motifs are unable to suppress any of the phenotypes exhibited by ymel deletion strains. Ymelp co-fractionates with proteins associated with the mitochondrial inner membrane, is tightly associated with this membrane, and is oriented with the bulk of the protein facing the matrix. Unassembled subunit II of cytochrome oxidase is stabilized in ymel yeast strains. The data support a model in which Ymelp is an ATP and zinc-dependent protease associated with the matrix side of the inner mitochondrial membrane. Subunit II of cytochrome oxidase, when not assembled into a higher order complex, is a likely substrate of Ymelp.

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“…Mutants were induced by ethidium bromide treatment essentially as described before (Slonimski et al, 1968). An overnight culture of strain X2180-1A and Y41 growing in synthetic medium (YNB) was inoculated into fresh YNB to a cell density of about 2 10 6 cells per ml.…”

Section: Construction Of Respiratory-deficient Mutantsmentioning

confidence: 99%

The Osmotic Hypersensitivity of the Yeast Saccharomyces cerevisiae is Strain and Growth Media Dependent: Quantitative Aspects of the Phenomenon

Blomberg

1997

Yeast

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Osmotic hypersensitivity is manifested as cellular death at magnitudes of osmotic stress that can support growth. Cellular capacity for survival when plated onto high NaCl media was examined for a number of laboratory and industrial strains of Saccharomyces cerevisiae. During respiro-fermentative growth in rich medium with glucose as energy and carbon source, the hypersensitivity phenomenon was fairly strain invariant with a threshold value of about 1 -NaCl; most strains fell within a 300 m range in LD 10 values (lethal dose yielding 10% survival). Furthermore, all but one of the strains displayed similar differential death responses above the threshold value, i.e. ten-fold decreased viability for every 250 m increase in salinity. Addition of small amounts of salt to the growth medium drastically improved tolerance and shifted the hypersensitivity threshold to higher NaCl concentrations. This salt-instigated tolerance could partly be reversed by washing in water. The washing procedure depleted cells of the glycerol that they had accumulated under saline growth, and the contribution from glycerol to the improved tolerance was about 50% in the two strains examined. Growth on derepressing carbon sources like galactose, ethanol or glycerol gave strain-dependent responses. The laboratory strain X2180-1A drastically improved tolerance while the bakers' yeast strain Y41 did so only marginally. It was concluded that all strains of S. cerevisiae display the osmotic hypersensitivity phenomenon in qualitative terms while the quantitative values differ. It was also proposed that growth rate does not dictate the level of osmotic hypersensitivity of S. cerevisiae.

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Ethidium bromide induced mutation of yeast mitochondria: Complete transformation of cells into respiratory deficient non-chromosomal “petites”

Cited by 489 publications

References 5 publications

In-vitro resistance to azoles associated with mitochondrial DNA deficiency in Candida glabrata

In-vitro resistance to azoles associated with mitochondrial DNA deficiency in Candida glabrata

Biochemical and functional analysis of the YME1 gene product, an ATP and zinc-dependent mitochondrial protease from S. cerevisiae.

The Osmotic Hypersensitivity of the Yeast Saccharomyces cerevisiae is Strain and Growth Media Dependent: Quantitative Aspects of the Phenomenon

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