Studies on Nuclear and Mitochondrial DNA of Saccharomyces cerevisiae

Dennis, E. S.; Goldthwaite, Claire; Zínker, Samuel; Lam, Ksl; Storm, Eva; Hirschberg, Rona; Blamire, John; Finkelstein, David; Marmur, Julius

doi:10.1101/sqb.1974.038.01.005

Cited by 28 publications

(11 citation statements)

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“…2). As expected (12,22), mitochondrial DNA was also synthesized during incubation in a-factor dsRNA (Fig. 3A).…”

Section: Resultssupporting

confidence: 79%

“…a-Factor is a yeast pheromone produced by cells of a mating type which arrests cells of a mating type late in the Gl phase of the cell cycle (9,23). The synthesis of RNA, protein, and mitochondrial DNA continues in a-factor treated cells (12,22,28), whereas synthesis of chromosomal and 2-,um DNAs is blocked (9,18 (Fig. 2).…”

Section: Resultsmentioning

confidence: 99%

“…To determine the ratios for M and L dsRNA's, the remaining sample was centrifuged to equilibrium in CsCl, and the dense regions of the gradients were pooled and then examined on agarose gels. The 3H/ 14C ratios for DNA reflect the cessation of chromosomal DNA replication (28) and the continued replication of mitochondrial DNA (12,22). The (20,35).…”

Section: Resultsmentioning

confidence: 99%

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Yeast L double-stranded ribonucleic acid is synthesized during the G1 phase but not the S phase of the cell cycle.

Zakian¹,

Wagner²,

Fangman³

1981

Mol. Cell. Biol.

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The cytoplasm of Saccharomyces cerevisiae contains two major classes of protein-encapsulated double-stranded ribonucleic acids (dsRNA's), L and M. Replication of L and M dsRNA's was examined in cells arrested in the G1 phase by either alpha-factor, a yeast mating pheromone, or the restrictive temperature for a cell cycle mutant (cdc7). [3H]uracil was added during the arrest periods to cells prelabeled with [14C]uracil, and replication was monitored by determining the ratio of 3H/14C for purified dsRNA's. Like mitochondrial deoxyribonucleic acid, both L and M dsRNA's were synthesized in the G1 arrested cells. The replication of L dsRNA was also examined during the S phase, using cells synchronized in two different ways. Cells containing the cdc7 mutation, treated sequentially with alpha-factor and then the restrictive temperature, enter a synchronous S phase when transferred to permissive temperature. When cells entered the S phase, synthesis of L dsRNA ceased, and little or no synthesis was detected throughout the S phase. Synthesis of L dsRNA was also observed in G1 phase cells isolated from asynchronous cultures by velocity centrifugation. Again, synthesis ceased when cells entered the S phase. These results indicate that L dsRNA replication is under cell cycle control. The control differs from that of mitochondrial deoxyribonucleic acid, which replicates in all phases of the cell cycle, and from that of 2-micron DNA, a multiple-copy plasmid whose replication is confined to the S phase.

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“…2). As expected (12,22), mitochondrial DNA was also synthesized during incubation in a-factor dsRNA (Fig. 3A).…”

Section: Resultssupporting

confidence: 79%

Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

See 1 more Smart Citation

Yeast L double-stranded ribonucleic acid is synthesized during the G1 phase but not the S phase of the cell cycle.

Zakian¹,

Wagner²,

Fangman³

1981

Mol. Cell. Biol.

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“…A diploid strain of Saccharomyces cerevisiae (Z193) was employed in this study. Z193 is heteroallelic at the lysine 2 (lys 2-1/lys 2-2) and tryptophan 5 (trp 5-R/trp [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] tubes and fractions were collected into 1 ml of 0.3 N NaOH containing 0.1 mg/ml of uracil. The fractions were hydrolyzed overnight at room temperature, refrigerated, neutralized with 0.5 ml of cold 0.6 N HCl and precipitated with 0.25 ml of cold…”

Section: Methodsmentioning

confidence: 99%

Single-strand scissions of chromosomal DNA during commitment to recombination at meiosis.

Jacobson¹,

Piñon²,

Esposito³

et al. 1975

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

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Genetic analysis of meiotic recombination has illustrated that recombination occurs by both gene conversion and reciprocal exchange (1). Molecular models proposed to explain these phenomena invoke the introduction of single-strand nicks (breakage of one phosphodiester bond) or gaps into the DNA of homologous chromosomes to initiate interactions which result in both conversion and reciprocal events (2, 3). The purpose of this study was to determine the timing and extent of single-strand and double-strand scissions of chromosomal DNA as diploid cells stimulated to enter meiosis become committed to genetic recombination. Yeast diploids are favorable material for a study of this type. Intact chromosomal DNA duplexes can be obtained by lysis of spheroplasts prepared from yeast cells. The chromosomes of yeast are unineme and consist of DNA duplexes ranging in molecular weight from 5 X 107 to 1.4 X 109 (4-6). These molecules can be fractionated by sedimentation velocity in neutral sucrose gradients under appropriate conditions (4, 5) and can be examined for single-strand nicks or gaps by analysis of sedimentation patterns in alkaline sucrose gradient (7).We have found that meiotic yeast cells do not accumulate double-strand breaks but do accumulate single-strand breaks after the onset of premeiotic DNA synthesis and commitment to genetic recombination. Nicking of both template and newly synthesized DNA was observed. When commitment to recombination is at the full meiotic level we estimate there are two to six single-strand scissions/109 daltons of DNA per 4C meiotic cell (4C indicates four copies of each locus). The relationship of these observations to related studies of meiotic DNA metabolism and pertinent yeast genetic data is discussed. MATERIALS AND METHODSOrganism. A diploid strain of Saccharomyces cerevisiae (Z193) was employed in this study. Z193 is heteroallelic at the lysine 2 (lys 2-1/lys 2-2) and tryptophan 5 (trp 5-R/trp [5][6][7][8][9][10][11][12][13][14][15][16][17][18][19][20] tubes and fractions were collected into 1 ml of 0.3 N NaOH containing 0.1 mg/ml of uracil. The fractions were hydrolyzed overnight at room temperature, refrigerated, neutralized with 0.5 ml of cold 0.6 N HCl and precipitated with 0.25 ml of cold

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“…Information concerning the physical properties, multiplicity and organization of ribosomal RNA cistrons in yeasts has been mainly derived from studies with Saccharomyces cerevisiae (Cryer et al, 1973;Kaback et al, 1973). These investigations have revealed that there are approximately 100 to 140 cistrons per haploid genome (the haploid genome size is approximately 5 x lo9 to 8 x lo9 daltons), coding for 25, 18, 5.8 and 5s rRNA each.…”

Section: Introductionmentioning

confidence: 99%

Ribosomal RNA Genes in Kluyveromyces marxianus

Ojha¹

1980

Microbiology

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449DNA from the yeast Kluyveromyces marxianus was studied for its heterogeneity and the multiplicity of rRNA cistrons. These cistrons banded at a slightly different density from the bulk DNA in preparative CsCl or Hg2+-CszSOa equilibrium density gradients. The reassociation kinetics of the bulk DNA showed that the repetitive fraction represented a small amount of the total cellular DNA (10%) and that the single copy fraction had a complexity of 6.3 x lo9 daltons. Approximately 2.2% of the DNA hybridized to 3H-labelled rRNA at saturation. On the basis of the above genome size, the multiplicity of the rRNA cistrons was calculated to be about 70 per haploid nucleus.

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Studies on Nuclear and Mitochondrial DNA of Saccharomyces cerevisiae

Cited by 28 publications

References 0 publications

Yeast L double-stranded ribonucleic acid is synthesized during the G1 phase but not the S phase of the cell cycle.

Yeast L double-stranded ribonucleic acid is synthesized during the G1 phase but not the S phase of the cell cycle.

Single-strand scissions of chromosomal DNA during commitment to recombination at meiosis.

Ribosomal RNA Genes in Kluyveromyces marxianus

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