Summary
Ribosomal RNA (rRNA) genes of Saccharomyces cerevisiae are clustered in a DNA “repeat unit” of 5.9 megadaltons with the gene order 5S-18S-5.8S-25S rRNA (Nath and Bollon, 1977). By using two restriction endonucleases, EcoRII and HindII, which generate DNA fragments that span contiguous portions of two “repeat units”, we report that the rRNA gene clusters are tandemly repeated without the intervention of additional spacer DNA.
The treatment of yeast DNA with the restriction endonucleases EcoRII and HindII result in the generation of 4 different DNA fragments that are of varying sizes and which hybridize with rRNA. The largest DNA fragments, 3.30 megadaltons in the case of HindII and 3.67 megadaltons in the case of EcoRII, encompass regions that code for the two opposite end regions of the 35S precuursor-rRNA. These two end regions are joined by a constant DNA segment of about 0.9 megadaltons in size of which a 0.08 megadalton segment codes for 5S rRNA. Since the 35S precursor-rRNA includes the 5.8S, 18S and 25S rRNA most of the “repeat units” containing the 4 rRNA coding genes in yeast are linked to each other contiguously without any intervening spacer DNA.
A composite map of the DNA restriction fragments obtained by the action of the restriction endonucleases EcoRI, EcoRII, HindII and HindIII on the 5.9 megadalton “repeat unit” is presented. Some striking features concerning the location of the restriction sites are noted. Of the total 17 DNA restriction sites present on each “repeat unit”, 9 are located at or near the 3 transcribed spacer regions contained in the 5 megadalton DNA segment that codes for the 35S precursor-rRNA. The 3 transcribed spacer regions in the 35S precursor-rRNA include the two external transcribed spacer regions and an internal transcribed spacer region, the latter representing the 5.8S rRNA.
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References
Bigger, C.H., Murray, K., Murray, N.E.: Recognition sequence of a restriction enzyme. Nature (Lond.) New Biol. 244, 7–10 (1973)
Bollon, A.P.: Regulation of the ilv 1 multifunctional gene in Saccharomyces cerevisiae. Molec. gen. Genet. 142, 1–12 (1975)
Boyer, H.W., Chow, L.T., Dugaiczyk, A., Hedgpeth, J., Goodman, H.M.: DNA substrate site for the EcoRII restriction endonuclease and modification methylase. Nature (Lond.) New Biol. 244, 40–43 (1973)
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)
Cramer, J.H., Farrelly, F.W., Barnitz, J.T., Rownd, R.H.: Construction and restriction endonuclease mapping of hybrid plasmids containing Saccharomyces cerevisiae ribosomal DNA. Molec. gen. Genet. 151, 229–244 (1977)
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)
Dawid, I.B., Brown, D.D., Reeder, R.H.: Composition and structure of chromosomal and amplified ribosomal DNA's of Xenopus laevis. J. molec. Biol. 51, 341–360 (1970)
Finkelstein, D.B., Blamire, J., Marmur, J.: Location of ribosomal RNA cistrons in yeast. Nature (Lond.) 240, 279–281 (1972)
Fukuhara, H.: Informational role of mitochondrial DNA studied by hybridization with different classes of RNA in yeast. Proc. nat. Acad. Sci. (Wash.) 58, 1065–1072 (1967)
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)
Kelly, T.J., Smith, H.O.: A restriction enzyme from Hemophilus influenza II. Base sequence of the recognition site. J. molec. Biol. 51, 393–409 (1970)
Maxam, A.M., Tizard, R., Skryabin, K.G., Gilbert, W.: Promoter region for yeast 5S ribosomal RNA. Nature (Lond.) 267, 643–645 (1977)
McLaughlin, C.S.: Yeast ribosomes: Genetics. In: Ribosomes (M. Nomura, A. Tissieres, and P. Lengyel, eds.), pp. 815–827. New York: Cold Spring Harbor Press 1974
Meyerink, J.H., Retél, J., Planta, R.J., Heidekamp, F.: Topographical analysis of yeast ribosomal DNA by cleavage with restriction endonucleases. Molec. biol. Reports (Holland) 2, 393–400 (1976)
Meyerink, J.H., Retél, J.: Topographical analysis of yeast ribosomal DNA by cleavage with restriction endonucleases. II. The physical map of EcroRI fragments. Nucl. Acid. Res. 3, 2697–2707 (1976)
Miyazaki, M.: Studies of the nucleotide sequence of pseudouridine containing 5S RNA from Saccharomyces cerevisiae. J. Biochem. (Japan) 75, 1407–1410 (1974)
Nath, K., Bollon, A.P.: Generation of discrete yeast DNA fragments by endonuclease R1. Nature (Lond.) 257, 155–157 (1975)
Nath, K., Bollon, A.P.: Characterization of yeast ribosomal DNA fragments generated by EcoRI restriction endonuclease. Molec. gen. Genet. 147, 153–168 (1976)
Nath, K., Bollon, A.P.: Organization of the yeast ribosomal RNA gene cluster via cloning and restriction analysis. J. biol. Chem. 252, 6562–6571 (1977)
Retél, J., Planta, R.J.: Nuclear satellite DNA's of yeast. Biochim. biophys. Acta (Amst.) 281, 299–309 (1972)
Retél, J., Van Keulen, H.: Characterization of yeast ribosomal DNA. Europ. J. Biochem. 58, 51–57 (1975)
Schweizer, E., Mackehnie, C., Halvorson, H.O.: The redundancy of ribosomal and transfer RNA genes in Saccharomyces cerevisiae. J. molec. Biol. 40, 261–277 (1969)
Seligy, V.L., James, A.P.: Multiplicity and distribution of rRNA cistrons among chromosome I and VII aneuploids of Saccharomyces cerevisiae. Exp. Cell Res. 105, 63–72 (1977)
Southern, E.M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. molec. Biol. 98, 503–517 (1975)
Udem, S.A., Warner, J.R.: Ribosomal RNA synthesis in Saccharomyces cerevisiae. J. molec. Biol. 65, 227–242 (1972)
Valenzuela, P., Bell, G.I., Masiarz, F.R., DeGennaro, L.J., Rutter, W.J.: Nucleotide sequence of the yeast 5S ribosomal RNA gene and adjacent putative control regions. Nature (Lond.) 267, 641–643 (1977)
Wellauer, P.K., Dawid, I.B., Brown, D.D., Reeder, R.H.: The molecular basis for length heterogeneity in ribosomal DNA from Xenopus laevis. J. molec. Biol. 105, 461–486 (1976)
Wellauer, P.K., Reeder, R.H., Dawid, I.B., Brown, D.D.: The arrangement of length heterogeneity in repeating units of amplified and chromosomal ribosomal DNA from Xenopus laevis. J. molec. Biol. 105, 487–505 (1976)
Wensink, P.C., Brown, D.D.: Denaturation map of the ribosomal DNA of Xenopus laevis. J. molec. Biol. 60, 235–247 (1971)
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Nath, K., Bollon, A.P. Restriction analysis of tandemly repeated yeast ribosomal RNA genes. Molec. Gen. Genet. 160, 235–245 (1978). https://doi.org/10.1007/BF00332967
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DOI: https://doi.org/10.1007/BF00332967