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Ribosomal DNA of the fly Sciara coprophila has a very small and homogeneous repeat unit

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Summary

In this report we show by hybridization of restriction fragments and by Miller spreads that the unit repeat of the fly Sciara coprophila is only 8.4 kb which is the smallest known for a multicellular eukaryote. The 8.4 kb EcoR1 fragment containing a complete unit of Sciara rDNA was cloned in pBR322, and mapped by the method of Parker (1977) and also by double digestion. The coding regions for 28S, 18S, and 5.8S RNA were localized by the method of Berk and Sharp (1977). From these data we conclude that the nontranscribed spacer, external transcribed spacer, and internal transcribed spacer are all shorter than in other organisms, thereby giving rise to the shorter overall rDNA repeat unit of Sciara.

At least 90% of the Sciara rDNA repeats are homogeneous, with a length of 8.4 kb, but a 700 bp ladder of minor bands can also be found in digestions of total genome DNA. This profile of major and minor bands is identical between the X and X′ chromosomes, as seen by a comparison of several genotypes.

There are only 45 rRNA genes per X chromosome of Sciara (Gerbi and Crouse, 1976). These can easily be counted by low magnification Miller speads which show that virtually all gene copies are actively being transcribed in the stage of spermatogenesis examined. This is the first demonstration for any reiterated gene family where all copies are shown to be simultaneously active.

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References

  • Barnett, T., Rae, P.M.M.: A 9.6 kb intervening sequence in D. virilis rDNA and sequence homology in rDNA interruptions of diverse species of Drosophila and other Diptera. (Cell, 1979 in press)

  • Beckingham, K., White, R.: Molecular characterization of ribosomal RNA genes of Calliphora erythrocephala. In: International Meeting on Molecular and Developmental Biology of Insects, Heraklion, Crete (in press, 1978).

  • Berk, A.J., Sharp, P.A.: Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell 12, 721–732 (1977)

    Google Scholar 

  • Bolivar, F., Rodriguez, R.L., Greene, P.J., Betlach, M.C., Heyneker, H.L., Boyer, H.W., Crosa, J.H., Falkow, S.: Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene 2, 95–113 (1977)

    Google Scholar 

  • Bos, J.L., Heyting, C., Borst, P., Arnberg, A.C., Van. Bruggen, E.F.J.: An insert in the single gene for the large ribosomal RNA in yeast mitochondrial DNA. Nature 275, 336–338 (1978)

    Google Scholar 

  • Botchan, P., Reeder, R.H., Dawid, I.B.: Restriction analysis of the nontranscribed spacers of Xenopus laevis ribosomal DNA. Cell 11, 599–607 (1977)

    Google Scholar 

  • Carson, H.L.: The selective elimination of inversion dicentric chromatids during meiosis in the eggs of Sciara impatiens. Genetics 31, 95–113 (1946)

    Google Scholar 

  • Clarke, L., Carbon, J.: Biochemical construction and selection of hybrid plasmids containing specific segments of the Escherichia coli genome. Proc. Natl. Acad. Sci. U.S.A. 72, 4361–4365 (1975)

    Google Scholar 

  • Clewell, D.B.: Nature of Col E1 plasmid replication in Escherichia coli in the presence of chloramphenicol. J. Bacteriol. 110, 667–676 (1972)

    Google Scholar 

  • Cockburn, A.F., Newkirk, M.J., Firtel, R.A.: Organization of the ribosomal RNA genes of Dictyostelium discoideum: mapping of the nontranscribed spacer regions. Cell 9, 605–613 (1976)

    Google Scholar 

  • Crouse, H.V.: X heterochromatin subdivision and cytogenetic analysis in Sciara coprophila (Diptera, Sciaridae). I. Centromere localization. Chromosoma 63, 39–55 (1977)

    Google Scholar 

  • Crouse, H.V., Gerbi, S.A., Liang, C.M., Magnus, L., Mercer, I.M.: Localization of ribosomal DNA within the proximal X heterochromatin of Sciara coprophila (Diptera, Sciaridae). Chromosoma 64, 305–318 (1977)

    Google Scholar 

  • Dawid, I.B., Long, E.: Ribosomal DNA and related sequences in Drosophila melanogaster. In: Alfred Benzon Symp. XIII. Copenhagen: Munksgaard, (in press, 1979)

    Google Scholar 

  • Dawid, I.B., Wellauer, P.K., Long, E.: Ribosomal DNA in Drosophila melanogaster. I. Isolation and characterization of cloned fragments. J. Mol. Biol. 126, 749–768 (1978)

    Google Scholar 

  • Denhardt, D.T.: A membrane-filter technique for the detection of complementary DNA. Biophys. Biochem. Res. Comm. 23, 641–646 (1966)

    Google Scholar 

  • DeVries, F.A.J., Collins, C.J., Jackson, D.A.: Joining of Simian Virus 40 DNA molecules of endonuclease R EcoR1 sites by polynucleotide ligase and analysis of the products by agarose gel electrophoresis. Biochim. Biophys. Acta 435, 213–227 (1976)

    Google Scholar 

  • Echalier, G., Ohanessian, A.: In vitro culture of Drosophila melanogaster embryonic cells. In Vitro 6, 162–172 (1970)

    Google Scholar 

  • Foe, V.E., Wilkinson, L.E., Laird, C.D.: Comparative organization of active transcription units in Oncopeltus fasciatus. Cell 9, 131–146 (1976)

    Google Scholar 

  • Gerbi, S.A.: Localization and characterisation of the ribosomal RNA cistrons in Sciara coprophila. J. Mol. Biol. 58, 499–511 (1971)

    Google Scholar 

  • Gerbi, S.A., Crouse, H.V.: Further studies on the ribosomal RNA cistrons of Sciara coprophila (Diptera). Genetics 83, 81–90 (1976)

    Google Scholar 

  • Glätzer, K.H.: Visualization of gene transcription in spermatocytes of Drosophila hydei. Chromosoma 53, 371–379 (1975)

    Google Scholar 

  • Glover, D.M., Hogness, D.S.: A novel arrangement of the 18S and 28S sequences in repeating unit of Drosophila melanogaster rDNA. Cell 10, 167–176 (1977)

    Google Scholar 

  • Grunstein, M., Hogness, D.S.: Colony hybridization: a method for the isolation of cloned DNAs that contain a specific gene. Proc. Natl. Acad. Sci. U.S.A. 72, 3961–3965 (1975)

    Google Scholar 

  • Helling, R.B., Goodman, H.M., Boyer, H.W.: Analysis of Endonuclease R. EcoR1 fragments of DNA from lambdoid bacteriophages and other viruses by agarose-gel electrophoresis. J. Virol. 14, 1235–1244 (1974)

    Google Scholar 

  • Hollenberg, C.P.: Proportionate representation of rDNA and Balbiani ring DNA in polytene chromosomes of Chironomus tentans. Chromosoma 57, 185–197 (1976)

    Google Scholar 

  • Jordan, B.R., Jourdan, R., Jacq, B.: Late steps in the maturation of Drosophila 26S ribosomal RNA: generation of 5.8S and 2S RNAs by cleavages occuring in the cytoplasm. J. Mol. Biol. 101, 85–105 (1976)

    Google Scholar 

  • Lambert, B., Egyhazi, E., Daneholt, B., Ringborg, U.: Quantitative micro-assay for RNA/DNA hybrids in the study of nucleolar RNA from Chironomus tentans salivary gland cells. Exp. Cell Res. 76, 369–380 (1973)

    Google Scholar 

  • Landy, A., Abelson, J., Goodman, H.M., Smith, J.D.: Specific hybridization of Tyrosine transfer ribonucleic acids with DNA from a transducing bacteriophage ϕ80 carrying the amber suppressor gene suIII. J. Mol. Biol. 29, 457–471 (1967)

    Google Scholar 

  • Maizels, N.: Dictyostelium 17S, 25S, and 5S rDNAs lie within a 38,000 base pair repeated unit. Cell 9, 431–438 (19760

    Google Scholar 

  • Mandel, M., Higa, A.: Calcium-dependent bacteriophage DNA infection. J. Mol. Biol. 53, 159–162 (1970)

    Google Scholar 

  • Maniatis, T., Jeffrey, A., Kleid, D.G.: Nucleotide sequence of the rightward operator of phage λ. Proc. Natl. Acad. Sci. U.S.A. 72, 1184–1188 (1975)

    Google Scholar 

  • Manning, R.F., Samols, D.R., Gage, L.P.: The genes for 18S, 5.8S and 28S ribosomal RNA of Bombyx mori are organized into tandem repeates of uniform length. Gene 4, 153–166 (1978)

    Google Scholar 

  • Marmur, J.: A procedure for the isolation of deoxyribonucleic acid from micro-organisms. J. Mol. Biol. 3, 208–218 (1961)

    Google Scholar 

  • McDonnell, M.W., Simon, M.N., Studies, F.W.: Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J. Mol. Biol. 110, 119–146 (1977)

    Google Scholar 

  • McKnight, S.L., Miller, O.L.: Ultrastructural patterns of RNA synthesis during early embryogenesis of Drosophila melanogaster. Cell 8, 305–319 (1976)

    Google Scholar 

  • Metz, C.W., Smith, H.B.: Further observations on the nature of the X-prime (X′) chromosome in Sciara. Proc. Natl. Acad. Sci. U.S.A. 17, 195–198 (1931)

    Google Scholar 

  • Meyer, G.F., Henning, W.: The nucleolus in primary spermatocytes of Drosophila hydei. Chromosoma 46, 121–144 (1974)

    Google Scholar 

  • Miller, O.L., Bakken, A.M.: Morphological studies of transcription. Acta Endocrinol. (Kbh.) [Suppl.] 168, 155–177 (1972)

    Google Scholar 

  • Miller, O.L., Beatty, B.R.: Visualization of nucleolar genes. Science 164, 955–957 (1969)

    Google Scholar 

  • Pardue, M.L., Gerbi, S.A., Eckhardt, R.A., Gall, J.G.: Cytological localization of DNA complementary to ribosomal RNA in polytene chromosomes of Diptera. Chromosoma 29, 268–290 (1970)

    Google Scholar 

  • Parker, R.C., Watson, R.M., Vinograd, J.: Mapping of closed circular DNAs by cleavage with restriction endonucleases and calibration by agarose gel electrophoresis. Proc. Natl. Acad. Sci. U.S.A. 74, 851–857 (1977)

    Google Scholar 

  • Peacock, A.C., Dingman, C.W.: Molecular weight estimation and separation of ribonucleic acid by electrophoresis in agaroseacrylamide composite gels. Biochemistry 7, 668–674 (1968)

    Google Scholar 

  • Pellegrini, M., Manning, J., Davidson, N.: Sequence arrangement of the rDNA of Drosophila melanogaster. Cell 10, 213–224 (1977)

    Google Scholar 

  • Pene, J.J., Knight, E., Darnell, J.E.: Characterization of a new low molecular weight RNA in HeLa ribosomes. J. Mol. Biol. 33, 609–623 (1968)

    Google Scholar 

  • Rae, P.M.M., Spear, B.B.: Macronuclear DNA of the hypotrichous ciliate Oxytricha fallax. Proc. Natl. Acad. Sci. U.S.A. 75, 4992–4996 (1978)

    Google Scholar 

  • Renkawitz, R., Kunz, W.: Independent replication of the ribosomal RNA genes of the polytrophic-meroistic ovaries of Calliphora erythrocephala, Drosophila hydei, and Sarcophaga barbata. Chromosoma 53, 131–140 (1975)

    Google Scholar 

  • Rochaix, J.D., Malnoe, P.: Anatomy of the chloroplast ribosomal DNA of Chlamydomonas reinhardii. Cell 15, 661–670 (1978)

    Google Scholar 

  • Scheer, U.: Changes of nucleosome frequency in nucleolar and nonnucleolar chromatin as a function of transcription: an electron microscope study. Cell 13, 535–549 (1978)

    Google Scholar 

  • Schneider, I.: Cell lines derived from late embryonic stages of Drosophila melanogaster. J. Embryol. Exp. Morphol. 27, 353–365 (1972)

    Google Scholar 

  • Shine, J., Dalgarno, L.: Occurence of heat-dissociable ribosomal RNA in insects: the presence of three polynucleotide chains in 26S RNA from cultured Aedes aegypti cells. J. Mol. Biol. 75, 57–72 (1973)

    Google Scholar 

  • Southern, E.M.: Detection of specific sequences among DNA fragments separated by gel electrophoresis. J. Mol. Biol. 93, 503–517 (1975)

    Google Scholar 

  • Steffan, A.: A generic revision of the family Sciaridae (Diptera) of America, north of Mexico. Univ. of California publication in entomology 44, 1–77 (1966).

    Google Scholar 

  • Suteliffe, J.G.: pBR322 restriction map derived from the DNA sequence: accurate DNA size markers up to 4361 nucleotide pairs long. Nucleic Acids Res. 5, 2721–2728 (1978)

    Google Scholar 

  • Tartof, K.D., Dawid, I.B.: Similarities and differences in the structure of X and Y chromosome rRNA genes in Drosophila. Nature 263, 27–30 (1978)

    Google Scholar 

  • Trendelenburg, M.F., Spring, H., Scheer, U., Franke, W.W.: Morphology of nucleolar cistrons in a plant cell, Acetabularia mediterranea. Proc. Natl. Acad. Sci. U.S.A. 71, 3626–3630 (1974)

    Google Scholar 

  • Wellauer, P.K., Dawid, I.B.: Secondary structure maps of ribosomal RNA and DNA. I. Processing of Xenopus laevis ribosomal RNA and structure of single-stranded ribosomal DNA. J. Mol. Biol. 89, 379–395 (1974)

    Google Scholar 

  • 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. Mol. Bio. 105, 461–486 (1976)

    Google Scholar 

  • Wellauer, P.K., Dawid, I.B.: The structural organization of ribosomal DNA in Drosophila melanogaster. Cell 10, 193–212 (1977)

    Google Scholar 

  • Wellauer, P.K., Dawid, I.B.: Ribosomal DNA in Drosophila melanogaster. II. Heteroduplex mapping of cloned and uncloned rDNA. J. Mol. Biol. 126, 769–782 (1978)

    Google Scholar 

  • Wellauer, P.K., Dawid, I.B., Tartof, K.D.: X and Y chromosomal ribosomal DNA of Drosophila: comparison of spacers and insertions. Cell 14, 269–278 (1978)

    Google Scholar 

  • White, R.L., Hogness, D.S.: R loop mapping of the 18S and 28S sequences in the long and short repeating units of Drosophila melanogaster rDNA. Cell 10, 177–192 (1977)

    Google Scholar 

  • Wild, M.A., Gall, J.G.: An intervening sequence in the gene coding for 25S ribosomal RNA of Tetrahymena pigmentosa. Cell 16, 565–573 (1979)

    Google Scholar 

  • Yao, M.-C., Gall, J.G.: A single integrated gene for ribosomal RNA in eucaryote, Tetrahymena pyriformis. Cell 12, 121–132 (1977)

    Google Scholar 

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Authors and Affiliations

  1. Division of Biology and Medicine, Brown University Providence, 92912, Rhode Island, U.S.A.

    Rainer Renkawitz & Susan A. Gerbi

  2. Institut für Allgemeine Biologie, Universität Düsseldorf, Universitätsstraße 1, D-4000, Düsseldorf, Federal Republic of Germany

    Karl Heinz Glätzer

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  1. Rainer Renkawitz
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  2. Susan A. Gerbi
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  3. Karl Heinz Glätzer
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Communicated by W. Gehring

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Renkawitz, R., Gerbi, S.A. & Glätzer, K.H. Ribosomal DNA of the fly Sciara coprophila has a very small and homogeneous repeat unit. Molec. Gen. Genet. 173, 1–13 (1979). https://doi.org/10.1007/BF00267685

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