Closed-Circular DNA from Mitochondrial-Enriched Fractions of Four Petite-Negative Yeasts

O'Connor, R M; McArthur, C. R.; Clark-Walker, G. D.

doi:10.1111/j.1432-1033.1975.tb04050.x

Cited by 47 publications

(20 citation statements)

References 29 publications

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“…In a separate study of circular DNA in another yeast, Torulopsis glabrata, we again found a class of non-mitochondrial molecules which had a length of 3.1 pm and a buoyant density of 1.709 g/cm3 (4). By analogy with the heavy satellite in S. cerevisiae it can be inferred that the 3.1 pm circular molecules-in this yeast code for ribosomal RNAs. Recently we have recommenced studying the 3 pm molecules because of their potential for mediating inter-species transfer of genes.…”

Section: Introductionmentioning

confidence: 85%

Hybridizable sequences between cytoplasmic ribosomal RNAs and 3 micron circular DNAs of Saccharomyces cerevisiae and Torulopsis glabrata

Cark-Walker

Azad

1980

Nucl Acids Res

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We have shown that 2.8 and 3.1 pm circular DNA molecules, previously reported to be present in Saccharomyces cerevisiae and Torulopsis glabrata respectively, contain sequences hybridizing to cytoplasmic ribosomal RNAs.In S. cerevisiae the 2.8 pm circular DNA appears to be identical to the rDNA repeating unit from nuclear DNA, both in length (approximately 9000 base pairs) and in the location of the 25, 18 and 5.8S rRNA sequences on the large HindIII fragment (6500 bp) and the presence of the 5S rRNA sequence on the small HindIII fragment. In S. cerevisiae the 4-5 copies of the 2.8 pm circular DNA molecules per cell, which have an extra-nuclear location, do not appear to be essential for cell viability as in one strain they were undetectable.

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Section: Introductionmentioning

confidence: 85%

Hybridizable sequences between cytoplasmic ribosomal RNAs and 3 micron circular DNAs of Saccharomyces cerevisiae and Torulopsis glabrata

Cark-Walker

Azad

1980

Nucl Acids Res

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“…Among different organisms, the mtDNA sizes are highly variable, ranging from the 5-Am circles of animals to the 30-Mm circles found in pea leaves (13,(34)(35)(36)(37)(38)(39)(40)(41)(42). However, this large variance in size does not necessarily imply that comparable differences in information content exist among these mtDNAs.…”

Section: Methodsmentioning

confidence: 99%

Isolation and characterization of mitochondrial DNA from Chlamydomonas reinhardtii.

Ryan¹,

Grant²,

Chiang³

et al. 1978

Proc. Natl. Acad. Sci. U.S.A.

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Mitochondrial DNA (mtDNA) has been isolated from a mitochondrial pellet of Chlamydomonas reinhardtii. The mtDNA has a buoyant density of 1.706 g/ml in CsCl, a melting temperature of 87.90 in standard saline citrate, and a nucleoside composition of 47.5% deoxyguanidine plus deoxycytidine with no odd nucleosides. Thermal denaturation and renaturation studies have shown that (i) mtDNA contains no extensive intramolecular heterogeneity nor significant base bias between the complementary polynucleotide chains and (ii) mtDNA renatures as a single homogeneous class with a kinetic complexity of 9.78 X 106 daltons. Although rare ( Primarily due to a number of advantageous biological characteristics, Chlamydomonas reinhardtia, a unicellular green alga, has been increasingly exploited for fruitful investigations of the cooperation between chloroplast and nuclear genomes in nucleic acid metabolism (1-4) and protein synthesis (5, 6). Genetic studies with a few non-Mendelian mutants have also suggested that there is probably another level of cooperation between the chloroplast and mitochondria genomes in organelle protein synthesis (7-9). Dissection of this postulated interorganelle cooperation at the molecular level should prove to be feasible and of interest, because C. reinhardtii is a facultative photosynthetic organism with an abundance of well-characterized nonphotosynthetic mutants (10). The unusually fragile structure of the single large chloroplast in C. reinhardtmi, however, has hampered repeated attempts in several laboratories to isolate contamination-free mitochondria for biochemical characterization.We report in this paper the successful isolation of a DNaseimpermeable mitochondrial preparation from a cell-wall-less mutant of C. reinhardtfil and the characterization of the mitochondrial DNA (mtDNA). We document here the basic physicochemical properties of this mtDNA and its complexity. The unexpected findings of our present study are that (i) the complexity of C. reinhardtii mtDNA is strikingly small in comparison with other plant mtDNAs but very close to that of animal mtDNA, and (ii) the reiterations of mtDNA and chloroplast DNA (cpDNA) molecules are comparable in the vegetative cells of C. reinhardtii. MATERIALS AND METHODSStrain and Culture Conditions. A mating type + subclone of the cell-wall-less mutant CW-15 (11) was used for all studies. Cells were grown in minimal medium supplemented with 0.2% sodium acetate (4). Twelve-or 18-liter cultures were illuminated with six circular 40-W cool-white fluorescent lamps and aerated with 3% CO2 in air.Mitochondria Isolation Procedures. Procedure A. Cells were collected at early stationary phase, washed once in buffer 1 (0.25 M sucrose/0.1 mM EDTA/10 mM Tris-HCl, pH 9), once in buffer 2 (0.25 M sucrose/0.1 mM EDTA/10 mM Tris-HCl, pH 7.4), and suspended in 10 ml of buffer 2 per liter of culture volume (12). This cell suspension was passed once through a chilled French pressure cell at 500 psi (34.5 X 102 kPa), diluted with an equal volume of buffer 2, and, af...

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“…According to established belief all known mitochondrial genomes in budding yeasts (with only two exceptions) consist of circular DNA molecules, which range in size from 19 kb in Candida (Torulopsis) glabrata, henceforth T.glabrata, to > 100 kb in Brettanomyces custersii (O'Connor et al, 1976;Hoeben and Clark-Walker, 1986). Copy number has been estimated to vary from 20 to 100 genome equivalents per cell, representing 5-20% of total cellular DNA (Dujon, 1981; Conrad and Newlon, 1982).…”

Section: Introductionmentioning

confidence: 99%

Rolling circle replication of DNA in yeast mitochondria.

1991

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The conformation of mitochondrial DNA (mtDNA) from yeasts has been examined by pulsed field gel electrophoresis and electron microscopy. The majority of mtDNA from Candida (Torulopsis) glabrata (mtDNA unit size, 19 kb) exists as linear molecules ranging in size from 50 to 150 kb or 2–7 genome units. A small proportion of mtDNA is present as supercoiled or relaxed circular molecules. Additional components, detected by electron microscopy, are circular molecules with either single‐ or double‐stranded tails (lariats). The presence of lariats, together with the observation that the majority of mtDNA is linear and 2–7 genome units in length, suggests that replication occurs by a rolling circle mechanism. Replication of mtDNA in other yeasts is thought to occur by the same mechanism. For Saccharomyces cerevisiae, the majority of mtDNA is linear and of heterogeneous length. Furthermore, linear DNA is the chief component of a plasmid, pMK2, when it is located in the mitochondrion of baker's yeast, although only circular DNA is detected when this plasmid occurs in the nucleus. The implications of long linear mtDNA for hypotheses concerning the ploidy paradox and the mechanism of the petite mutation are discussed.

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Closed-Circular DNA from Mitochondrial-Enriched Fractions of Four Petite-Negative Yeasts

Cited by 47 publications

References 29 publications

Hybridizable sequences between cytoplasmic ribosomal RNAs and 3 micron circular DNAs of Saccharomyces cerevisiae and Torulopsis glabrata

Hybridizable sequences between cytoplasmic ribosomal RNAs and 3 micron circular DNAs of Saccharomyces cerevisiae and Torulopsis glabrata

Isolation and characterization of mitochondrial DNA from Chlamydomonas reinhardtii.

Rolling circle replication of DNA in yeast mitochondria.

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