Summary
Mitochondrial transfer RNA genes have been ordered relative to the position of five mitochondrial drug resistance markers, namely, chloramphenicol (C), erythromycin (E), oligomycin I and II (OI, OII), and paromomycin (P). Forty-six petite yeast clones that were genetically characterized with respect to these markers were used for a study of these relationships. Different regions of the mitochondrial genome are deleted in these individual mutants, resulting in variable loss of genetic markers. Mitochondrial DNA was isolated from each mutant strain and hybridized with eleven individual mitochondrial transfer RNAs. The following results were obtained: i) Of the seven petite clones that retained C, E, and P resistance markers (but not OI or OII), four carried all eleven transfer RNA genes examined; the other three clones lost several transfer RNA genes, probably by secondary internal deletion; ii) Prolyl and valyl transfer RNA genes were located close to the P marker, whereas the histidyl transfer RNA gene was close to the C marker; iii) Except for a glutamyl transfer RNA gene that was loosely associated with the OI region, no other transfer RNA genes were found in petite clones retaining only the OI and/or the OII markers; and iv) Two distinct mitochondrial genes were found for glutamyl transfer RNA, they were not homologous in DNA sequence and were located at two separate loci.
The data indicate that the petite mitochondrial genome is the result of a primary deletion followed by successive additional deletions. Thus an unequivocal gene arrangement cannot be readily established by deletion mapping with petite mutants alone. Nevertheless, we have derived a tentative circular map of the yeast mitochondrial genome from the data; the map indicates that all but one of the transfer RNA genes are found between the C and P markers without forming a tight cluster. The following arrangement is suggested:-P-pro-val-ile-(phe, ala, tyr, asp)-glu2-(lys-leu)-his-C-E-OI-glu1-OII-P-.
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Avner, P.R., Coen, D., Dujon, B., Slonimski, P.P.: Mitochondrial genetics. IV. Allelism and mapping studies of oligomycin resistant mutants in S. cerevisiae. Molec. gen. Genet 125, 9–52 (1973)
Casey, J., Cohen, M., Rabinowitz, M., Fukuhara, H., Getz, G. S., Fukuhara, H.: Hybridization of mitochondrial transfer RNA's with mitochondrial and nuclear DNA of grande (wild type) yeast. J. molec. Biol. 63, 431–440 (1972)
Casey, J., Gordon, P., Rabinowitz, M.: Characterication of mitochondrial DNA from grande and petite yeasts by renaturation and denaturation analysis and by transfer RNA hybridization; evidence for internal repetition or heterogeneity in mitochondrial DNA populations. Biochemistry 13, 1059–1067 (1974)
Casey, J., Hsu, H. U., Rabinowitz, M., Getz, G. S., Fukuhara, H.: Transfer RNA genes in the mitochondrial DNA of cytoplasmic petite mutants of Saccharomyces cerevisiae. J. molec. Biol. 88, 717–733 (1974)
Cohen, M., Rabinowitz, M.: Analysis of grande and petite yeast mitochondrial DNA by tRNA hybridization. Biochim. biophys. Acta (Amst.) 281, 192–201 (1972)
Deutsch, J., Dujon, B., Netter, P., Petrochilo, E., Slonimski, P. P., Bolotin-Fukuhara, M., Coen, D.: Mitochondrial genetics VI. The petite mutantion in Saccharomyces cerevisiae: Interrelations between the loss of the ρ+ factor and the loss of the drug resistance mitochondrial genetic markers. Genetics 76, 195–219 (1974)
Faye, G., Fukuhara, H., Grandchamp, C., Lazowska, J., Michel, F., Casey, J., Getz, G. S., Locker, J., Rabinowitz, M., Bolotin-Fukuhara, M., Coen, D., Deutsch, J., Dujon, B., Netter, P., Slonimski, P. P.: Mitochondrial nucleic acids in the petite colonie mutants: deletions and repetitions of genes. Biochimie 55, 779–792 (1973)
Faye, G., Kujawa, C., Dujon, B., Bolotin-Fukuhara, M., Wolf, K., Fukuhara, H., Slonimski, P. P.: Localization of the gene coding for the 16S ribosomal mitochondrial RNA using rho− mutants of Saccharomyces cerevisiae. J. molec. Biol. 99, 203–217 (1975)
Faye, G., Kujawa, C., Fukuhara, H.: Physical and genetic organization of petite and grande yeast mitochondrial IV. In vivo transcription products of mitochondrial DNA snd localization of 23S ribosomal RNA in petite mutants of Saccharomyces cerevisiae. J. molec. Biol. 88, 185–203 (1974)
Fukuhara, H., Faye, G., Michel, F., Lazowska, J., Deutsch, J., Bolotin-Fukuhara, M., Slonimski, P. P.: Physical and genetic organization of petite and grande yeast mitochondrial DNA. I. Studies by RNA-DNA hybridization. Molec. gen. Genet. 130, 215–238 (1974)
Hollenberg, C. P., Borst, P., Van Bruggen, E. F. J.: Mitochondrial DNA V. A 25 μ closed circular duplex DNA molecule in wild-type yeast mitochondria. Structure and genetic complexity. Biochem. biophys. Acta (Amst.) 209, 1–15 (1970)
Lazowska, J., Michel, F., Faye, G., Fukuhara, H., Slonimski, P. P.: Physical and genetic organization of petite and grande yeast mitochondrial DNA. II. DNA-DNA hybridization studies and buoyant density determinations. J. molec. Biol. 85, 393–410 (1974)
Locker, J., Rabinowitz, M., Getz, G. S.: Electron microscopic and renaturation kinetic analysis of mitochondrial DNA of cytoplasmic petite mutants of Saccharomyces cerevisiae. J. molec. Biol. 88, 489–502 (1974)
Locker, J., Rabinowitz, M., Getz, G. S.: Tandem inverted repeats in mitochondrial DNA of petite mutants of Saccharomyces cerevisiae. Proc. nat. Acad. Sci. (Wash.) 71, 1366–1370 (1974)
Martin, N., Rabinowitz, M., Fukuhara, H.: Isoaccepting mitochondrial glutamyl tRNA species transcribed from different regions of the mitochondrial genome of Saccharomyces cerevisiae. J. molec. Biol. (in press) (1976)
Michel, F., Lazowska, J., Faye, G., Fukuhara, H., Slonimski, P. P.: Physical and genetic organization of petite and grande yeast mitochondrial DNA III. High resolution melting and reassociation studies. J. molec. Biol. 85, 411–431 (1974)
Netter, P., Petrochilo, E., Slonimski, P. P., Bolotin-Fukuhara, M., Coen, D., Deutsch, J., Dujon, P.: Mitochondrial genetics VII. Allelism and mapping studies of ribosomal mutants resistant to chloramphenicol erythromycin and spiramycin in S. cerevisiae. Genetics 78, 1063–1100 (1974)
Plischke, M., Von Vorstel, R. C., Mortimer, E. K., Cohn, W. E.: In: The handbook of biochemistry and molecular biology (Fasman, G. C., ed.), 3rd ed. Cleveland: Chemical Rubber Co. Press (in press)
Wetmur, J. G., Davidson, N.: Kinetics of renaturation of DNA. J. molec. Biol. 31, 349–370 (1968)
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Communicated by F.Kaudewitz
Supported in part by Cancer Center CCRC 111B-3. Present address: Laboratoire de Biologie Generale, Universite Paris-Sud Orsay, 91405, France
The Franklin McLean Memorial Research Institute is operated by the University of Chicago for the U.S. Energy Research and Development Administration under Contract E(11-1)69
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Fukuhara, H., Bolotin-Fukuhara, M., Hsu, HJ. et al. Deletion mapping of mitochondrial transfer RNA genes in Saccharomyces cerevisiae by means of cytoplasmic petite mutants. Molec. gen. Genet. 145, 7–17 (1976). https://doi.org/10.1007/BF00331551
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DOI: https://doi.org/10.1007/BF00331551