Physical and genetic organization of petite and grande yeast mitochondrial DNA

Michel, François; Lazowska, Jaga; Faye, Gérard; Fukuhara, Hiroshi; Słonimski, Piotr P.

doi:10.1016/0022-2836(74)90441-0

Cited by 51 publications

(11 citation statements)

References 26 publications

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“…Double strands of DNA are melted to single strands during the process of temperature rising. Considerable evidence has been presented by various investigators (1)(2)(3)(4)(5)(6)(7)(8), demonstrating that the melting of DNA has been detected as multimodal curves of the change in optical density as a function of temperature. It has been suggested that the melting process of DNA takes place blockwise within the molecule depending on the intramolecular heterogeneity of base pair distribution.…”

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confidence: 99%

Differential scanning calorimetric study of plasmid DNA.

Maeda

Kawai

Fujita

et al. 1984

J. Gen. Appl. Microbiol.

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ANDDifferential scanning calorimetry (DSC) of double stranded DNA was carried out using plasmid pJL3-TB5 DNA (5277 base pairs), whose entire nucleotide sequence has been determined. The DSC curve for the linear DNA of the plasmid used in the present study gave seven separate peaks including two main peaks which range from 82° to 98°, suggesting that DNA melts independently at several cooperative blocks along the chain. The DSC curve obtained agreed with the histogram of the frequency versus GC content which was constructed from the GC distribution map along the DNA chain of pJL3-TB5. These results are obvious evidence that the thermal transition of DNA occurs blockwise along the molecular chain. The two main peaks were estimated to be due to the heat transition of the two distinct regions, containing 46-50 % and 54-58 % GC, respectively. DSC of covalently closed circular (ccc-) DNA of the same plasmid did not show any heat transition peak, indicating that ccc-form is stabilized compared with linear form. Double strands of DNA are melted to single strands during the process of temperature rising. Considerable evidence has been presented by various investigators (1-8), demonstrating that the melting of DNA has been detected as multimodal curves of the change in optical density as a function of temperature. It has been suggested that the melting process of DNA takes place blockwise within the molecule depending on the intramolecular heterogeneity of base pair distribution. However, it has been difficult to make a detailed assignment of a melting profile to the sequences of DNA, since the DNA samples used are large and their exact nucleotide sequences have not been available. A number of investigations on the melting of DNA have been also performed by a calorimetric method, since this phenomenon is accompanied by heat absorption. However, distinct evidence 289

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confidence: 99%

Differential scanning calorimetric study of plasmid DNA.

Maeda

Kawai

Fujita

et al. 1984

J. Gen. Appl. Microbiol.

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“…It is now well documented that petites are the result of extensive deletions of wild-type mtDNA sequences (13-s15) and, moreover, that individual petite isolates can retain any part of the mitochondrial genome (16, 17), most probably in a geneamplified configuration (18)(19)(20) (22) and mit-loci (11). What is intrinsically uncertain in such analyses, however, is whether or not the particular allele localized by the phenotype measured represents the structural gene sequence for a specified polypeptide product.…”

Section: Introductionmentioning

confidence: 99%

Expression of petite mitochondrial DNA in vivo: zygotic gene rescue.

Strausberg¹,

Butow²

1977

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

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A protocol is introduced for probing the organization and regulation of expression of the yeast mitochondrial genome, termed "zygotic gene rescue." The procedure is based on the notion that genes retained on mitochondrial DNA of petites can be expressed in zygotes of a cross between petite and wild type. To test the validity of this notion, we have taken advantage of our ability to discriminate, by mobility differences on sodium dodecyl sulfate/polyacrylamide gels, different forms of the product of alleles of the mitochondrial gene, varl. In petite strains that have retained the varl gene, its characteristic product appears in zygotes 4-5 hr after mating; no product is observed in petite strains deleted in the varl locus. Our studies indicate that (i) expression in the zygote of the varl gene in the petite genome is not exclusively the result of recombination with mitochondrial DNA of the wild-type tester, and (ii) the var) gene is probably reiterated in the petite mitochondrial genome. The strength of the technique of zygotic gene rescue in the analysis of the mitochondrial genome is discussed.Recent advances in genetic and physical mapping of the yeast mitochondrial genome have provided considerable information on the identification and localization of a number of mitochondrial genes. Among these are included drug resistance loci (1-3), genes specifying rRNAs (4-6) and tRNAs (7-9), and a variety of loci termed mit (10, 11). The latter, when mutated, lead to an inability of the strain to grow on nonfermentable carbon sources as a result of specific alterations in cytochrome oxidase, the cytochrome be1 complex, or the oligomycin-sensitive ATPase complex (12). These complexes are composed of nuclear encoded, cytoplasmically synthesized subunits and subunits that are translated on mitochondrial ribosomes and generally believed to be encoded by mitochondrial DNA (mtDNA). It is of obvious interest to know the precise location of sequences on the yeast mitochondrial genome corresponding to the structural genes for these and other, less well characterized gene products of mtDNA.In this communication we introduce a procedure for probing the organization of the yeast mitochondrial genome. This procedure, termed "zygotic gene rescue," is based on the premise that the mtDNA of yeast cells carrying the vegetative petite mutation (p-) is senseful and could, in principle, express structural gene sequences in tlvo given a functional transcriptional and translational apparatus.It is now well documented that petites are the result of extensive deletions of wild-type mtDNA sequences (13-s15) and, moreover, that individual petite isolates can retain any part of the mitochondrial genome (16, 17), most probably in a geneamplified configuration (18)(19)(20) (22) and mit-loci (11). What is intrinsically uncertain in such analyses, however, is whether or not the particular allele localized by the phenotype measured represents the structural gene sequence for a specified polypeptide product. We reasoned that it might be possibl...

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“…Renaturation kinetic analysis (4,5) and DNA-DNA hybridization studies (6)(7)(8)(9) indicate that petite mtDNAs contain deletions of 20% to more than 99% of the grande (wild-type) mitochondrial genome. By electron microscopic examination, a tandem repeat arrangement of petite mtDNA has been demonstrated, and in some cases there are inverted repeats of the mitochondrial genome, i.e., palindromes (2,3,10).…”

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confidence: 99%

“…Mutations 0144 and 0145 come from Dr. P. Avner, P454 from Dr. K. Wolf, and others from the C. G. M. stocks. The full genotypes of the p+ strains, from which p-clones were isolated, are the following: MH41-7B, a ade2 his1 CR321 ER514 OR145 PR454; MH32-12D, a ade2 his1 CR321 ER221 OR144 PR454; TR3-15A, a try2 his1 CR321 ER514 OR1 pR454-* The second-order rate constant of reassociation of the mtDNA (k2) was determined by a procedure similar to that described by Michel et al (4), and is expressed as the ratio of the k2 to that of grande mtDNA. For most studies, mtDNA was prepared by CsCl centrifugation after lysis of mitochondria, washed five times, which were prepared from protoplasts as described (5,16).…”

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confidence: 99%

Restriction endonuclease analysis of mitochondrial DNA from grande and genetically characterized cytoplasmic petite clones of Saccharomyces cerevisiae.

Morimoto¹,

Lewin²,

Hsu³

et al. 1975

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

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Digestion of grande mitochondrial DNA (mtDNA) by EcoRI restriction endonuclease gives rise to nine fragments with a total molecular weight of 51.8 X 106. HindIII digestion yields six fragments with a similar total molecular weight. Specific restriction fragments can be detected despite the fact that yeast mtDNA consists of a heterogeneous distribution of randomly broken molecules. Digestion patterns of 10 genetically characterized petite clones containing various combinations of five antiobiotic resistance markers indicate that the petite mtDNA predominantly represents deletion of the grande genome. The petite mtDNAs contained up to seven EcoRI restriction fragments which comigrate with grande restriction fragments, and at least one fragment that did not correspond to any in the grande. Some strains contained multiple fragments with mobility different from that of grande; these fragments were usually present in less than molar concentrations. The genetic markers were associated with individual sets of restriction fragments. However, several internal inconsistencies prevent the construction of a definitive genetic fragment map. These anomalies, together with the digestion patterns, provide strong evidence that, in addition to single contiguous deletion, other changes such as multiple deletion and heterogeneity of mtDNA populations are present in some of the petite mtDNAs.It is now well established that mitochondrial DNA (mtDNA) of cytoplasmic petite mutants is greatly altered (1-3). Renaturation kinetic analysis (4, 5) and DNA-DNA hybridization studies (6-9) indicate that petite mtDNAs contain deletions of 20% to more than 99% of the grande (wild-type) mitochondrial genome. By electron microscopic examination, a tandem repeat arrangement of petite mtDNA has been demonstrated, and in some cases there are inverted repeats of the mitochondrial genome, i.e., palindromes (2, 3, 10). Although deletion represents the most prominent alteration, other changes may be present in some strains, including heterogeneity of mtDNA populations (10-12), amplification of some segments of the mtDNA (10-12), and base sequence change (6, 9).Based on the assumption that deletion is the predominant alteration in petite mtDNA, our laboratories have attempted to map mitochondrial tRNA and rRNA genes by using a series of petite mutants. These studies have provided a partial ordering of the tRNA cistrons (11, §

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Physical and genetic organization of petite and grande yeast mitochondrial DNA

Cited by 51 publications

References 26 publications

Differential scanning calorimetric study of plasmid DNA.

Differential scanning calorimetric study of plasmid DNA.

Expression of petite mitochondrial DNA in vivo: zygotic gene rescue.

Restriction endonuclease analysis of mitochondrial DNA from grande and genetically characterized cytoplasmic petite clones of Saccharomyces cerevisiae.

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