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Regulation of the expression of iso 2-cytochrome c gene in S. cerevisiae: cloning of the positive regulatory gene CYP1 and identification of the region of its target sequence on the structural gene CYP3

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Summary

CYP1 is a trans acting regulatory locus modulating both iso 1- and iso 2-cytochrome c synthesis. Genetical analysis of various mutated alleles has allowed us to identify the gene product as a positive regulatory element.

The region of the target sequence of the CYP1 product on the iso 2-cytochrome c structural gene was located by molecular and genetic analysis of two cis acting mutations located at the CYP3 locus: CYP3-36 and CYP3-4, which have been shown to arise from the integration of TY1 elements near the promoter site. Determination of the amount of iso 2-cytochrome c synthesized by strains bearing various genetic constructions, in which the cis acting mutations were associated with different alleles of the CYP1 trans acting locus, showed that TY1 inserted into CYP3-36 extinguishes the activation function due to a mutated overproducer allele CYP1-18, while CYP3-4 amplifies this function. This result identifies at least a part of the target sequence of the CYP1 product within the region separating the two TY1 insertions.

To clone the CYP1 gene, we took advantage of the iso 2-cytochrome c overproducer phenotype of the mutated allele CYP1-18, which confers a Lactate+ phenotype on an iso 1-cytochrome c-deficient strain. Such a phenotype allowed the isolation of a recombinant plasmid YEpJFM1 carrying the mutated allele, able to complement on lactate medium a lactate - recipient strain. The identity of the YEpJFM1 sequence with the chromosomal gene was confirmed by homologous recombination at the CYP1 locus.

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Authors and Affiliations

  1. Centre de Génétique Moléculaire du C.N.R.S. laboratoire propre associé à l'Université P et M. Curie, F-91190, Gif-sur-Yvette, France

    Jacqueline Verdière & Francine Creusot

  2. Laboratoire de Biologie Génétique et Moléculaire, Université Paris XI, F-91405, Orsay, France

    Michel Guérineau

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  1. Jacqueline Verdière
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  2. Francine Creusot
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  3. Michel Guérineau
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Communicated by W. Gajewski

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Verdière, J., Creusot, F. & Guérineau, M. Regulation of the expression of iso 2-cytochrome c gene in S. cerevisiae: cloning of the positive regulatory gene CYP1 and identification of the region of its target sequence on the structural gene CYP3 . Mol Gen Genet 199, 524–533 (1985). https://doi.org/10.1007/BF00330769

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  • DOI: https://doi.org/10.1007/BF00330769

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