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  • 1
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: The Saccharomyces cerevisiae nuclear gene OXA1, which is conserved from prokaryotes to human, was shown to be essential for cytochrome c oxidase and F1F0–ATP synthase biogenesis. We have searched for an orthologue of OXA1 in Schizosaccharomyces pombe, another yeast that is highly diverged from S. cerevisiae and which could more closely model higher eukaryotes. In particular, S. pombe exhibits a limited growth under anaerobic conditions and is petite negative, that is it does not tolerate large deletions of its mitochondrial DNA. Surprisingly, two S. pombe cDNAs able to complement an S. cerevisiae oxa1 mutation were isolated. The corresponding genes have different chromosomal locations and intron contents. They encode distinct proteins, both sharing a weak sequence identity one with the other and with Oxa1p. A phenotypic analysis of both single inactivations demonstrates that only one gene is essential for respiration in S. pombe. However, the double inactivation is lethal. This work gives new insight into the dependence of S. pombe viability upon oxa1 function, providing evidence of a connection between petite negativity, a functional respiratory chain and F1F0–ATP synthase complex in S. pombe.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1365-2958
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Most cellular ATP is produced within the mitochondria from ADP and Pi which are delivered across the inner-membrane by specific nuclearly encoded polytopic carriers. In Saccharomyces cerevisiae, some of these carriers and in particular the ADP/ATP carrier, are represented by several related isoforms that are distinct in their pattern of expression. Until now, only one mitochondrial Pi carrier (mPic) form, encoded by the MIR1 gene in S. cerevisiae, has been described. Here we show that the gene product encoded by the YER053C ORF also participates in the delivery of phosphate to the mitochondria. We have called this gene PIC2 for Pi carrier isoform 2. Overexpression of PIC2 compensates for the mitochondrial defect of the double mutant Δmir1 Δpic2 and restores phosphate transport activity in mitochondria swelling experiments. The existence of two isoforms of mPic does not seem to be restricted to S. cerevisiae as two Arabidopsis thaliana cDNAs encoding two different mPic-like proteins are also able to complement the double mutant Δmir1 Δpic2. Finally, we demonstrate that Pic2p is a mitochondrial protein and that its steady state level increases at high temperature. We propose that Pic2p is a minor form of mPic which plays a role under specific stress conditions.
    Type of Medium: Electronic Resource
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  • 3
    Electronic Resource
    Electronic Resource
    [s.l.] : Nature Publishing Group
    Nature 298 (1982), S. 628-632 
    ISSN: 1476-4687
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
    Notes: [Auszug] An extragenic suppressor mutation, mim2–1, which compensates yeast mitochondrial mutants deficient in splicing of the cytochrome b gene, has been mapped and sequenced. The mutation is due to a single G → A transition in the long open reading frame of the fourth intron of the oxidase ...
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1432-0983
    Keywords: Key words Mitochondria ; Membrane protein ; Respiratory complex assembly
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract We have previously isolated the yeast nuclear gene OXA1 and showed that Oxa1p is required for the formation of the cytochrome c oxidase and ATP synthase complexes. We have expressed Oxa1p in E. coli and shown that it is toxic and rapidly degraded. Nevertheless, a truncated protein was successfully expressed and antibodies have been raised against this truncated protein. These antibodies recognise a protein in mitochondrially enriched fractions. In vitro mitochondrial import experiments demonstrate that the import of Oxa1p is accompanied by the cleavage of a long pre-sequence. Osmotic swelling and alkaline carbonate extraction show that Oxa1p is an integral membrane protein located in the inner membrane of mitochondria. The relationships between the sub-mitochondrial location and the function of Oxa1p are discussed.
    Type of Medium: Electronic Resource
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  • 5
    ISSN: 1432-0983
    Keywords: Yeast mitochondrial ; Paromomycin ; Phenotypic suppression ; Cytoplasmic element [psi]
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Paromomycin has been found to suppress certain nonsense mutations located in several mitochondrial genes in yeast. In the mosaic genes, paromomycin preferentially suppresses those mutations located in the introns. There is a strong correlation between this phenotypic suppression by paromomycin and the genetic suppression due to various informational mitoribosomal suppressors. No effect of the cytoplasmic element [psi] on mitoribosomal protein synthesis was observed. This work provides strong evidence for the translation of mRNA maturase proteins from open reading frames of the mitochondrial introns.
    Type of Medium: Electronic Resource
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  • 6
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary The effects of hydroxyurea, an inhibitor of DNA synthesis, on the transmission and recombination of mitochondrial genes conferring resistance to different antibiotics in Saccharomyces cerevisiae have been studied. Under the conditions used hydroxyurea does not induce respiratory deficient mutants in treated cultures. Homosexual and heterosexual crosses have been performed in which one of the parents was grown for several generations in a medium containing 10-1 M hydroxyurea prior to mating and the other parent was untreated. In all cases, the transmission of mitochondrial alleles originating from the hydroxyurea treated parent increased. In homosexual crosses the increase of transmission is coordinated for all the alleles. In heterosexual crosses, there is a differential increase of transmission of the different alleles depending on the distance of the marker considered from ω. All the observed effects can be easily interpreted according to the predictions of the model proposed by Dujon et al. (1974), if one assumes that hydroxyurea treatment increases the input of mitochondrial alleles of the treated parent without major modifications of the frequency of recombination. For each cross the relative copy number of mitDNA of the treated parent has been calculated and the increase so found is in good agreement with the modification of the overall frequency of recombinants observed. Effects of hydroxyurea on input can be interpreted if one assumes that hydroxyurea has a differential effect on mitochondrial and nuclear DNA synthesis: nuclear DNA synthesis would be inhibited while mitochondrial DNA synthesis would still continue. In conclusion, we propose a new powerful genetic tool for the study of the transmission and recombination of mitochondrial genes which offers the possibility of experimentally controlling input, without inducing rho- mutation.
    Type of Medium: Electronic Resource
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  • 7
    Electronic Resource
    Electronic Resource
    Springer
    Molecular genetics and genomics 167 (1979), S. 301-308 
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Fifty eight mitochondrial mutants (p + mit- mutants), all deficient in cytochrome oxidase activity and previously assigned to the genetic region oxi3 on the mitochondrial DNA, were mapped by the method of “petite deletion mapping”. This procedure resulted in the identification of at least twenty one different classes of oxi3 mutants, which could be arranged in a linear order. Moreover, it provided a set of twenty three p - petite mutants, each containing a differentially deleted mit DNA segment included in the oxi3 region. The two sets of mutants, p + oxi3 - and p - oxi3 +, will be of interest for a further genetic and physical analysis of this mitochondrial DNA segment which spans over about ten thousand base pairs and controls the subunit I of cytochrome oxidase.
    Type of Medium: Electronic Resource
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  • 8
    ISSN: 1617-4623
    Keywords: Yeast ; Nuclear genes ; Mitochondrial translation ; Mitochondrial splicing ; Suppression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We have cloned three distinct nuclear genes, NAM1, NAM7, and NAM8, which alleviate mitochondrial intron mutations of the cytochrome b and COXI (subunit I of cytochrome oxidase) genes when present on multicopy plasmids. These nuclear genes show no sequence homology to each other and are localized on different chromosomes: NAM1 on chromosome IV, NAM7 on chromosome XIII and NAM8 on chromosome VIII. Sequence analysis of the NAM1 gene shows that it encodes a protein of 440 amino acids with a typical presequence that would target the protein to the mitochondrial matrix. Inactivation of the NAM1 gene by gene transplacement leads to a dramatic reduction of the overall synthesis of mitochondrial protein, and a complete absence of the COXI protein which is the result of a specific block in COXI pre-mRNA splicing. The possible mechanisms by which the NAM1 gene product may function are discussed.
    Type of Medium: Electronic Resource
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  • 9
    ISSN: 1617-4623
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A series of mutants called ebi, less inducible by ethidium bromide than the parental strain for the ρ+→ρ− mutation have been isolated after E.M.S. mutagenesis. Some of the ebi mutants also show an important accumulation of ρ− cells, in the absence of ethidium bromide. Ebi mutations are nuclearly inherited as shown by meiotic segregation. The effects of these mutants on the transmission and recombination of mitochondrial genes among the diploid progeny of crosses have been studied. Some of the ebi mutants show a non coordinated transmission of the oli1 mitochondrial marker with respect to other mitochondrial markers unexpected for homosexual crosses. This bias which is independent from ω will be discussed in relation to the segregation and recombination. No significant decrease of the frequency of recombinants has been detected.
    Type of Medium: Electronic Resource
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  • 10
    ISSN: 1617-4623
    Keywords: Yeast ; Mitochondria ; pre-mRNA splicing ; tRNA synthetase gene ; Incipient evolution
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary We studied the NAM2 genes of Saccharomyces douglasii and Saccharomyces cerevisiae, and showed that they are interchangeable for all the known functions of these genes, both mitochondrial protein synthesis and mitochondrial mRNA splicing. This confirms the prediction that the S. douglasii NAM2D gene encodes the mitochondrial leucyl tRNA synthetase (EC 6.1.1.4). The observation that these enzymes are interchangeable for their mRNA splicing functions, even though there are significant differences in the intron/exon structure of their mitochondrial genome, suggests that they may have a general role in yeast mitochondrial RNA splicing. A short open reading frame (ORF) precedes the synthetase-encoding ORF, and we showed that at least in S. cerevisiae this is not essential for the expression of the gene; however, it may be involved in a more subtle type of regulation. Sequence comparisons of S. douglasii and S. cerevisiae revealed a particularly interesting situation from the evolutionary point of view. It appears that the two yeasts have diverged relatively recently: there is remarkable nucleotide sequence conservation, with no deletions or insertions, but numerous (albeit non-saturating) silent substitutions resulting from transitions. This applies not only to the NAM2 coding regions, but also to two other ORFs flanking the NAM2 ORF. The regions between the ORFs (believed to be intergenic regions) are much less conserved, with several deletions and insertions. Thus S. douglasii and S. cerevisiae provide an ideal system for the study of molecular evolution, being two yeasts “caught in the act” of speciation.
    Type of Medium: Electronic Resource
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