Summary
The size and the degree of homogeneity of the repetitive units in purified ribosomal DNA (γ DNA) from Saccharomyces cerevisiae have been analyzed by restriction endonuclease digestion and heteroduplex mapping. Digestion of the γ DNA with EcoRI yields seven fragments, digestion with Hind II+III yields five fragments, digestion with Hind III alone yields two fragments, and digestion with Sma I yields one fragment. The sum of the fragment molecular weights after digestion with each of the endonucleases is 5.5–5.6×106. When the DNA strands of the Sma I fragment are dissociated and reannealed, only homoduplexes are formed. We have concluded from these results that the repeating units in yeast ribosomal DNA are 5.6×106 daltons and are homogeneous in size and composition.
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Aarstad, K., Øyen, T.B.: On the distribution of 5S RNA cistrons on the genome of Saccharomyces cerevisiae. FEBS Letters 51, 227–231 (1975)
Berger, S., Schweiger, H.G.: An apparent lack of nontranscribed spacers in rDNA of a green alga. Molec. gen. Genet. 139, 269–275 (1975)
Bhargava, M.M., Cramer, J.H., Halvorson, H.O.: Isolation of high molecular weight DNA from yeast nuclei. Analyt. Biochem. 49, 276–284 (1972)
Corneo, G., Moore, C., Sanadi, D.R., Grossman, L.I., Marmur, J.: Mitochondrial DNA in yeast and some mammalian species. Science 151, 687–689 (1966)
Cramer, J.H., Bhargava, M.M., Halvorson, H.O.: Isolation and characterization of γ DNA of Saccharomyces cerevisiae. J. molec. Biol. 71, 11–20 (1972)
Davis, R.W., Simon, M., Davidson, N.: Electron microscope heteroduplex methods for mapping regions of base sequence homology in nucleic acids. In: Methods in enzymology (L. Grossman and K. Moldave, eds.), Vol. 21D, pp. 413–428. New York: Academic Press, Inc. 1971
Finkelstein, D.B., Blamire, J., Marmur, J.: Location of ribosomal RNA cistrons in yeast. Nature (Lond.) New Biol. 240, 279–281 (1972)
Freifelder, D.: Molecular weights of coliphages and coliphage DNA. J. molec. Biol. 54, 567–577 (1970)
Hedgpeth, J., Goodman, H.M., Boyer, H.W.: DNA nucleotide sequence restricted by the RI endonuclease. Proc. nat. Acad. Sci. (Wash.) 69, 3448–3452 (1972)
Inman, R.B., Schnös, M.: Partial denaturation of thymine- and 5-bromouracil-containing λ DNA in alkali. J. molec. Biol. 49, 93–98 (1970)
Kaback, D.B., Bhargava, M.M., Halvorson, H.O.: Location and arrangement of genes coding for ribosomal RNA in Saccharomyces cerevisiae. J. molec. Biol. 79, 735–739 (1973)
Karijord, Ø., Øyen, T.B.: Study of a haploid yeast strain with an unusually high rDNA content. II. Fractionation of the DNA in preparative Ag+-Cs2SO4 density gradients. Biochim. biophys. Acta (Amst.) 383, 255–265 (1975)
McLaughlin, C.S.: Yeast ribosomes: Genetics. In: Ribosomes (M. Nomura, A. Tissières, and P. Lengyel, eds.), pp. 815–827. Cold Spring Harbor Laboratory (1974)
Moustacchi, E., Williamson, D.H.: Physiological variations in satellite components of yeast DNA detected by density gradient centrifugation. Biochem. biophys. Res. Commun. 23, 56–61 (1966)
Nath, K., Bollon, A.P.: Generation of discrete yeast DNA fragments by endonuclease RI. Nature (Lond.) 257, 155–157 (1975)
Old, R., Murray, K., Roizes, G.: Recognition sequence of restriction endonuclease III from Hemophilus influenzae. J. molec. Biol. 92, 331–339 (1975)
Rubin, G.M., Sulston, J.E.: Physical linkage of the 5S cistrons to the 18S and 28S ribosomal RNA cistrons in Saccharomyces cerevisiae. J. molec. Biol. 79, 521–530 (1973)
Schweizer, E., MacKechnie, C., Halvorson, H.O.: The redundancy of ribosomal and transfer RNA genes in Saccharomyces cerevisiae. J. molec. Biol. 40, 261–277 (1969)
Sharp, P.A., Sugden, B., Sambrook, J.: Detection of two restriction endonuclease activities in Haemophilus parainfluenzae using analytical agarose-ethidium bromide electrophoresis. Biochemistry 12, 3055–3063 (1973)
Tanaka, N., Cramer, J.H., Rownd, R.H.: EcoRI restriction endonuclease map of the composite R plasmid NRl. J. Bact. 127, 619–636 (1976)
Tanaka, T., Weisblum, B.: Construction of a colicin El-R factor composite plasmid in vitro: Means for amplification of deoxyribonucleic acid. J. Bact. 121, 354–362 (1975)
Tewari, K.K., Jayaraman, J., Mahler, H.R.: Separation and characterization of mitochondrial DNA from yeast. Biochem. biophys. Res. Commun 21, 141–148 (1965)
Udem, S.A., Warner, J.R.: Ribosomal RNA synthesis in Saccharomyces cerevisiae. J. molec. Biol. 65, 227–242 (1972)
Younghusband, H.B., Egan, J.B., Inman, R.B.: Characterization of the DNA from bacteriophage P2-186. Hybrids and physical mapping of the 186 chromosome. Molec. gen. Genet. 140, 101–110 (1975)
Wellauer, P.K., Dawid, I.B.: Secondary structure maps of ribosomal RNA and DNA I. Processing of Xenopus laevis ribosomal lec.Biol. 89, 379–395 (1974)
Wellauer, P.K., Reeder, R.H.: A comparison of the structural organization of amplified ribosomal DNA from Xenopus mulleri and Xenopus laevis. J. molec. Biol. 94, 151–161 (1975)
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Communicated by F. Kaudewitz
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Cramer, J.H., Farrelly, F.W. & Rownd, R.H. Restriction endonuclease analysis of ribosomal DNA from Saccharomyces cerevisiae . Molec. Gen. Genet. 148, 233–241 (1976). https://doi.org/10.1007/BF00332897
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DOI: https://doi.org/10.1007/BF00332897