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Length heterogeneity of the rRNA precursor in cucumber (Cucumis sativus)

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Abstract

The length homogeneous part of the intergenic spacer (IGS) of the 18S–25S ribosomal RNA genes of cucumber (Cucumis sativus) was characterized by sequencing 2389 bp preceding the 18S rRNA coding region of a 12.5 kbp repeat type. This part of the IGS is composed of repeated elements and shows a very complex structural organization. Most obvious is a 119 bp element which is repeated seven times. A single transcription initiation site (TIS) was detected by a ‘T4 polymerase stop’ experiment upstream of these repetitions giving rise to a 2013 bp 5′ external transcribed spacer (ETS) for cucumber. Nuclease mapping showed several transcription termination sites (TTS): the first one is located 350 bp downstream of the 25S rRNA coding region, the others are found within the duplications of this region accounting for the length heterogeneity of cucumber rDNA [7]. Therefore, the TTS is repeated two or three times in the IGS depending on the length of the respective repeat classes and the rRNA precursor is heterogeneous in length varying from approx. 8000 to 11000 nts.

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References

  1. Appels R, Moran LB, Gustafson JP: The structure of DNA from rye (Secale cereale) NOR R1 locus and its behaviour in wheat backgrounds. Can J Genet Cytol 28: 673–685 (1986).

    Google Scholar 

  2. Barker RF, Harberd NP, Jarvis MG, Flavell RB: Structure and evolution of the intergenic region in a ribosomal DNA repeat unit of wheat. J Mol Biol 201: 1–17 (1988).

    PubMed  Google Scholar 

  3. Birnboim HC: A rapid alkaline extraction method for the isolation of plasmid DNA. Meth Enzymol 100: 243–255 (1983).

    PubMed  Google Scholar 

  4. Delcasso-Tremousaygue D, Grellet F, Panabieres F, Ananiev ED, Delseny M: Structural and transcriptional characterization of the external spacer of a ribosomal RNA nuclear gene from higher plant. Eur J Biochem 172: 767–776 (1988).

    PubMed  Google Scholar 

  5. Flavell RB, O'Dell M, Thompson WT, Vincentz M, Sardana R, Barker RF. The differential expression of ribosomal RNA genes. Phil Trans R Soc Lond B314: 385–397 (1986).

    Google Scholar 

  6. Ganal M, Hemleben V: Comparison of the ribosomal RNA genes in four closely related Cucurbitaceae. Pl Syst Evol 154: 63–77 (1986).

    Google Scholar 

  7. Ganal M, Torres R, Hemleben V: Complex structure of the ribosomal DNA spacer of Cucumis sativus (cucumber). Mol Gen Genet 212: 548–554 (1988).

    Article  PubMed  Google Scholar 

  8. Gerstner J, Schiebel K, vonWaldburg G, Hemleben V: Complex organization of the length heterogeneous 5′ external spacer of mung bean (Vigna radiata) ribosomal DNA. Genome 30: 723–733 (1988).

    PubMed  Google Scholar 

  9. Grellet F, Delcasso-Tremousaygue D, Delseny M: Isolation and characterization of an unusual repeated sequence from the ribosomal intergenic spacer of the crucifer Sisymbrium irio. Plant Mol Biol 12: 695–706 (1989).

    Google Scholar 

  10. Gruendler P, Unfried I, Pointner R, Schweizer D: Nucleotide sequence of the 25S–18S ribosomal gene spacer from Arabidopsis thaliana. Nucleic Acids Res 17: 6395–6396 (1989).

    PubMed  Google Scholar 

  11. Grummt I, Maier U, Öhrlein A, Hassouna N, Bachellerie J: Transcription of mouse rDNA terminates downstream of the 3′ end of the 25S RNA and involves interaction of factors with repeated sequences in the 3′ spacer. Cell 43: 801–810 (1985).

    Article  PubMed  Google Scholar 

  12. Grummt I, Kuhn A, Bartsch I, Rosenbauer H: A transcription terminator upstream of the mouse rDNA initiation site affects rRNA synthesis. Cell 47: 901–911 (1986).

    Article  PubMed  Google Scholar 

  13. Hemleben V, Ganal M, Gerstner J, Schiebel K, Torres RA: Organization and length heterogeneity of plant ribosomal RNA genes. In: Kahl G (ed) The Architecture of Eukaryotic Genes. VHC, Weinheim (1988).

    Google Scholar 

  14. Kavanagh T, Timmis J: Heterogeneity in cucumber ribosomal DNA. Theor Appl Genet 72: 337–345 (1986).

    Article  Google Scholar 

  15. Kelly RJ, Siegel A: The Cucurbita maxima ribosomal DNA intergenic spacer has a complex structure. Gene 80: 239–248 (1989).

    Article  PubMed  Google Scholar 

  16. Klösgen RB, Gierl A, Schwarz-Sommer Z, Saedler H: Molecular analysis of the waxy locus of Zea mays. Mol Gen Genet 203: 237–244 (1986).

    Article  Google Scholar 

  17. Labhardt P, Reeder RH: Characterization of three sites of RNA 3′ end formation in the Xenopus ribosomal gene spacer. Cell 45: 431–443 (1986).

    Article  PubMed  Google Scholar 

  18. Long EO, Dawid IB: Repeated genes in eukaryotes. Ann Rev Biochem 49: 727–764 (1980).

    Article  PubMed  Google Scholar 

  19. Mandal RK: The organisation and transcription of eukaryotic ribosomal RNA genes. Progr Nucleic Acids Res 31: 115–160 (1984).

    Google Scholar 

  20. Maniatis T, Fritsch EF, Sambrook J: Molecular Cloning: A Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY (1982).

    Google Scholar 

  21. McMullen MD, Hunter B, Phillips RL, Rubenstein J: The structure of the maize ribosomal DNA spacer region. Nucleic Acids Res 14: 4953–4968 (1986).

    PubMed  Google Scholar 

  22. McStay B, Reeder RH: A termination site for Xenopus RNA polymerase I also acts as an element of an adjacent promoter. Cell 47: 913–920 (1986).

    Article  PubMed  Google Scholar 

  23. Messing J, Vieira J: A new pair of M13 vectors either DNA strand or double-digest restriction fragments. Gene 19: 269–276 (1983).

    Article  Google Scholar 

  24. Mitchelson K, Moss T: The enhancement of ribosomal transcription by the recycling of RNA polymerase I. Nucl Acids Res 15: 9577–9596 (1987).

    PubMed  Google Scholar 

  25. Moss T: A transcriptional function for the repetitive ribosomal spacer in Xenopus laevis. Nature 302: 223–228 (1983).

    PubMed  Google Scholar 

  26. Pustell JM, Katafos FC: A convenient and adaptable package of computer programs for DNA and protein sequence management, analysis and homology determination. Nucleic Acids Res 12: 643–655 (1984).

    PubMed  Google Scholar 

  27. Reeder RH: Enhancers and ribosomal gene spacers. Cell 38: 349–351 (1984).

    Article  PubMed  Google Scholar 

  28. Rogers SO, Bendich AJ: Ribosomal RNA genes in plants: variability in copy number and in the intergenic spacer. Plant Mol Biol 9: 509–520 (1987).

    Google Scholar 

  29. Sanger F, Nicklen S, Coulson A: DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci USA 74: 5463–5467 (1977).

    PubMed  Google Scholar 

  30. Schiebel K, vonWaldburg G, Gerstner J, Hemleben V: Termination of transcription of ribosomal RNA genes of mung bean occurs within a 175 bp repetitive element of the spacer region. Mol Gen Genet 218: 302–307 (1989).

    Google Scholar 

  31. Schmidt-Puchta W, Günther I, Sänger HL: Nucleotide sequence of the intergenic spacer (IGS) of the tomato ribosomal DNA. Plant Mol Biol 13: 251–253 (1989).

    Article  PubMed  Google Scholar 

  32. Schmitz ML, Maier UG, Brown JWS, Feix G: Specific binding of nuclear proteins to the promoter region of a maize nuclear rRNA gene unit. J Biol Chem 264: 1467–1472 (1989).

    PubMed  Google Scholar 

  33. Sollner-Webb B, Tower J: Transcription of cloned eukaryotic ribosomal RNA genes. Ann Rev Biochem 55: 801–830 (1986).

    Article  PubMed  Google Scholar 

  34. Taira T, Kato K, Tanifuji S: Difference between two major size classes of carrot rDNA repeating units is due to reiteration of sequences of about 460 bp in the large spacer. Mol Gen Genet 213: 170–174 (1988).

    Article  PubMed  Google Scholar 

  35. Toloczyki C, Feix G: Occurrence of 9 homologous repeat units in the exteral spacer region of a nuclear maize rRNA gene unit. Nucleic Acids Res 14: 4969–4986 (1986).

    PubMed  Google Scholar 

  36. Torres RA, Zentgraf U, Hemleben V: Species and genus specificity of the intergenic spacer (IGS) in the ribosomal RNA genes of Cucurbitaceae. Z Naturforsch 44c: 1029–1034 (1989).

    Google Scholar 

  37. Torres RA, Ganal M, Hemleben V: GC balance in the internal transcribed spacers ITS 1 and ITS 2 of nuclear ribosomal genes. J Mol Evol, in press (1990).

  38. Vincentz M, Flavell RB: Mapping of ribosomal RNA transcripts in wheat. Plant Cell 1: 579–589 (1989).

    Article  PubMed  Google Scholar 

  39. Yanisch-Perron C, Vieira J, Messing J: Improved M13 phage cloning vectors and host strains: Nucleotide sequences of M13 mp 18 und pUC 19 vectors. Gene 33: 103–119 (1985).

    Article  PubMed  Google Scholar 

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  1. M. Ganal

    Present address: Department of Plant Breeding and Biometry, Cornell University, 416 Bradfield Hall, 14853, Ithaca, NY, USA

Authors and Affiliations

  1. Lehrstuhl für Allgemeine Genetik, Biologisches Institut der Universität Tübingen, Auf der Morgenstelle 28, D-7400, Tübingen, FRG

    U. Zentgraf, M. Ganal & V. Hemleben

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  1. U. Zentgraf
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  2. M. Ganal
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  3. V. Hemleben
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Zentgraf, U., Ganal, M. & Hemleben, V. Length heterogeneity of the rRNA precursor in cucumber (Cucumis sativus). Plant Mol Biol 15, 465–474 (1990). https://doi.org/10.1007/BF00019163

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  • DOI: https://doi.org/10.1007/BF00019163

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