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Physical mapping of genes on yeast mitochondrial DNA: Localization of antibiotic resistance loci, and rRNA and tRNA genes

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We have physically mapped the loci conferring resistance to antibiotics that inhibit mitochondrial protein synthesis (erythromycin, chloramphenicol and paromomycin) or respiration (oligomycin I and II), as well as the 21s and 14s rRNA and tRNA genes on the restriction map of the mitochondrial genome of the yeast Saccharomyces cerevisiae. The mitochondrial genes were localized by hybridization of labeled RNA probes to restriction fragments of grande (strain MH41-7B) mitochondrial DNA (mtDNA)1 generated by endonucleases EcoRI, HpaI, BamHI, HindIII, SalI, PstI and HhaI. We have derived the HhaI restriction fragment map of MH41-7B mit DNA, to be added to our previously reported maps for the six other endonucleases.

The antibiotic resistance loci (ant R) were mapped by hybridization of 3H-cRNA transcribed from single marker petite mtDNA's of low kinetic complexity to grande restriction fragments. We have chosen the single Sal I site as the origin of the circular physical map and have positioned the antibiotic loci as follows: C (99.5-1.Ou)-P(27-36.Ou)-OII (58.3-62u)-OI (80-84u)-E (94.4-98.4u). The 21s rRNA is localized at 94.4-99.2u, and the 14s rRNA is positioned between 36.2-39.8u. The two rRNA species are separated by 36% of the genome. Total mitochondrial tRNA labeled with 125I hybridized primarily to two regions of the genome, at 99.5-11.5u and 34-44u. A third region of hybridization was occasionally detected at 70-76u, which probably corresponds to seryl and glutamyl tRNA genes, previously located to this region by petite deletion mapping.

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Author notes

  1. Nancy C. Martin (Postdoctoral Fellow of the American Cancer Society)

    Present address: Department of Biochemistry, University of Minnesota School of Medicine, Minneapolis, Minnesota

Authors and Affiliations

  1. Department of Biology, The University of Chicago Pritzker School of Medicine, Chicago, Illinois, USA

    Richard Morimoto, Sylvie Merten, Alfred Lewin, Nancy C. Martin (Postdoctoral Fellow of the American Cancer Society) & Murray Rabinowitz

  2. Department of Medicine, The University of Chicago Pritzker School of Medicine, Chicago, Illinois, USA

    Richard Morimoto, Sylvie Merten, Alfred Lewin, Nancy C. Martin (Postdoctoral Fellow of the American Cancer Society) & Murray Rabinowitz

  3. Department of Biochemistry, The University of Chicago Pritzker School of Medicine, Chicago, Illinois, USA

    Richard Morimoto, Sylvie Merten, Alfred Lewin, Nancy C. Martin (Postdoctoral Fellow of the American Cancer Society) & Murray Rabinowitz

  4. Franklin McLean Memorial Research Institute, Chicago, Illinois, USA

    Richard Morimoto, Sylvie Merten, Alfred Lewin, Nancy C. Martin (Postdoctoral Fellow of the American Cancer Society) & Murray Rabinowitz

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  1. Richard Morimoto
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Additional information

Communicated by F. Kaudewitz

Supported by USPHS Training Grant T32-GM-07197.

Supported by USPHS Training Grant 5-T01-GM-0090-19.

The Franklin McLean Memorial Research Institute is operated by the University of Chicago for the U. S. Energy Research and Development Administration under Contract EY-76-C-02-0069.

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Morimoto, R., Merten, S., Lewin, A. et al. Physical mapping of genes on yeast mitochondrial DNA: Localization of antibiotic resistance loci, and rRNA and tRNA genes. Molec. Gen. Genet. 163, 241–255 (1978). https://doi.org/10.1007/BF00271954

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