Skip to main content
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1984 Jan 25;12(2):959–972. doi: 10.1093/nar/12.2.959

Nucleotide sequence of the 5'-terminal coding region for pre-rRNA and mature 17S rRNA in Tetrahymena thermophila rDNA.

J Engberg, N Din, H Saiga, T Higashinakagawa
PMCID: PMC318548  PMID: 6320127

Abstract

The 5'-terminus of 35S pre-rRNA and mature 17S rRNA of Tetrahymena thermophila was mapped on cloned rDNA fragments by S1 nuclease protection experiments. A single site for transcription initiation was observed when pre-rRNA prepared by three different methods was used as RNA probe. These mapping results were unambiguously confirmed by sequencing the 5'-terminal region of in vitro capped 35S pre-rRNA. DNA sequence analysis of about 520 nucleotides upstream of the transcription initiation site revealed several distinct sets of highly conserved repeat sequences. In addition, the 840 nucleotides downstream of the transcription initiation site (+ 1) was determined and shown to include the 5'-terminus of the 17S rRNA coding region at position + 647. A region surrounding the position + 195 contains an inverted repeat sequence which could be the structural basis for the recently described premature transcription termination event in this organism (Kister et al. (1983) Nucl. Acids Res. 11, 3487-3502).

Full text

PDF
959

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bayev A. A., Georgiev O. I., Hadjiolov A. A., Kermekchiev M. B., Nikolaev N., Skryabin K. G., Zakharyev V. M. The structure of the yeast ribosomal RNA genes. 2. The nucleotide sequence of the initiation site for ribosomal RNA transcription. Nucleic Acids Res. 1980 Nov 11;8(21):4919–4926. doi: 10.1093/nar/8.21.4919. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Benoist C., Chambon P. In vivo sequence requirements of the SV40 early promotor region. Nature. 1981 Mar 26;290(5804):304–310. doi: 10.1038/290304a0. [DOI] [PubMed] [Google Scholar]
  3. Bogenhagen D. F., Sakonju S., Brown D. D. A control region in the center of the 5S RNA gene directs specific initiation of transcription: II. The 3' border of the region. Cell. 1980 Jan;19(1):27–35. doi: 10.1016/0092-8674(80)90385-2. [DOI] [PubMed] [Google Scholar]
  4. Cech T. R., Rio D. C. Localization of transcribed regions on extrachromosomal ribosomal RNA genes of Tetrahymena thermophila by R-loop mapping. Proc Natl Acad Sci U S A. 1979 Oct;76(10):5051–5055. doi: 10.1073/pnas.76.10.5051. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Darnell J. E., Jr Variety in the level of gene control in eukaryotic cells. Nature. 1982 Jun 3;297(5865):365–371. doi: 10.1038/297365a0. [DOI] [PubMed] [Google Scholar]
  6. DeFranco D., Schmidt O., Söll D. Two control regions for eukaryotic tRNA gene transcription. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3365–3368. doi: 10.1073/pnas.77.6.3365. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dierks P., van Ooyen A., Mantei N., Weissmann C. DNA sequences preceding the rabbit beta-globin gene are required for formation in mouse L cells of beta-globin RNA with the correct 5' terminus. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1411–1415. doi: 10.1073/pnas.78.3.1411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Donis-Keller H., Maxam A. M., Gilbert W. Mapping adenines, guanines, and pyrimidines in RNA. Nucleic Acids Res. 1977 Aug;4(8):2527–2538. doi: 10.1093/nar/4.8.2527. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Engberg J., Nasir-ud-Din, Eckert W. A., Kaffenberger W., Pearlman R. E. Detailed transcription map of the extrachromosomal ribosomal RNA genes in Tetrahymena thermophila. J Mol Biol. 1980 Sep 25;142(3):289–313. doi: 10.1016/0022-2836(80)90274-0. [DOI] [PubMed] [Google Scholar]
  10. Financsek I., Mizumoto K., Muramatsu M. Nucleotide sequence of the transcription initiation region of a rat ribosomal RNA gene. Gene. 1982 May;18(2):115–122. doi: 10.1016/0378-1119(82)90109-3. [DOI] [PubMed] [Google Scholar]
  11. Fowlkes D. M., Shenk T. Transcriptional control regions of the adenovirus VAI RNA gene. Cell. 1980 Nov;22(2 Pt 2):405–413. doi: 10.1016/0092-8674(80)90351-7. [DOI] [PubMed] [Google Scholar]
  12. Gabrielsen O. S., Oyen T. B. Yeast RNA polymerase I binds preferentially to A+T-rich linkers in rDNA. Nucleic Acids Res. 1982 Oct 11;10(19):5893–5904. doi: 10.1093/nar/10.19.5893. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Georgiev O. I., Nikolaev N., Hadjiolov A. A., Skryabin K. G., Zakharyev V. M., Bayev A. A. The structure of the yeast ribosomal RNA genes. 4. Complete sequence of the 25 S rRNA gene from Saccharomyces cerevisae. Nucleic Acids Res. 1981 Dec 21;9(24):6953–6958. doi: 10.1093/nar/9.24.6953. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Grosschedl R., Birnstiel M. L. Identification of regulatory sequences in the prelude sequences of an H2A histone gene by the study of specific deletion mutants in vivo. Proc Natl Acad Sci U S A. 1980 Mar;77(3):1432–1436. doi: 10.1073/pnas.77.3.1432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Grummt I. Mapping of a mouse ribosomal DNA promoter by in vitro transcription. Nucleic Acids Res. 1981 Nov 25;9(22):6093–6102. doi: 10.1093/nar/9.22.6093. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Gruss P., Dhar R., Khoury G. Simian virus 40 tandem repeated sequences as an element of the early promoter. Proc Natl Acad Sci U S A. 1981 Feb;78(2):943–947. doi: 10.1073/pnas.78.2.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Higashinakagawa T., Saiga H., Shintani N., Narushima-Iio M., Mita T. Localization of putative transcription initiation site on the cloned rDNA fragment of Tetrahymena pyriformis. Nucleic Acids Res. 1981 Nov 25;9(22):5905–5916. doi: 10.1093/nar/9.22.5905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Hoshikawa Y., Iida Y., Iwabuchi M. Nucleotide sequence of the transcriptional initiation region of Dictyostelium discoideum rRNA gene and comparison of the initiation regions of three lower eukaryotes' genes. Nucleic Acids Res. 1983 Mar 25;11(6):1725–1734. doi: 10.1093/nar/11.6.1725. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kiss G. B., Amin A. A., Pearlman R. E. Two separate regions of the extrachromosomal ribosomal deoxyribonucleic acid of Tetrahymena thermophila enable autonomous replication of plasmids in Saccharomyces cerevisiae. Mol Cell Biol. 1981 Jun;1(6):535–543. doi: 10.1128/mcb.1.6.535. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kister K. P., Müller B., Eckert W. A. Complex endonucleolytic cleavage pattern during early events in the processing of pre-rRNA in the lower eukaryote, Tetrahymena thermophila. Nucleic Acids Res. 1983 Jun 11;11(11):3487–3502. doi: 10.1093/nar/11.11.3487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Long E. O., Rebbert M. L., Dawid I. B. Nucleotide sequence of the initiation site for ribosomal RNA transcription in Drosophila melanogaster: comparison of genes with and without insertions. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1513–1517. doi: 10.1073/pnas.78.3.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lykkesfeldt A. E., Andersen H. A. Preferential inhibition of rDNA transcription by 5-bromodeoxyuridine. J Cell Sci. 1977 Jun;25:95–102. doi: 10.1242/jcs.25.1.95. [DOI] [PubMed] [Google Scholar]
  23. Maden B. E., Moss M., Salim M. Nucleotide sequence of an external transcribed spacer in Xenopus laevis rDNA: sequences flanking the 5' and 3' ends of 18S rRNA are non-complementary. Nucleic Acids Res. 1982 Apr 10;10(7):2387–2398. doi: 10.1093/nar/10.7.2387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Mathis D. J., Chambon P. The SV40 early region TATA box is required for accurate in vitro initiation of transcription. Nature. 1981 Mar 26;290(5804):310–315. doi: 10.1038/290310a0. [DOI] [PubMed] [Google Scholar]
  25. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  26. Miller K. G., Sollner-Webb B. Transcription of mouse rRNA genes by RNA polymerase I: in vitro and in vivo initiation and processing sites. Cell. 1981 Nov;27(1 Pt 2):165–174. doi: 10.1016/0092-8674(81)90370-6. [DOI] [PubMed] [Google Scholar]
  27. Moss T. A transcriptional function for the repetitive ribosomal spacer in Xenopus laevis. Nature. 1983 Mar 17;302(5905):223–228. doi: 10.1038/302223a0. [DOI] [PubMed] [Google Scholar]
  28. Niles E. G. 5' -and 3'-terminal nucleotide sequences of Tetrahymena pyriformis 17S rRNA. Biochemistry. 1977 May 31;16(11):2380–2383. doi: 10.1021/bi00630a011. [DOI] [PubMed] [Google Scholar]
  29. Niles E. G. Isolation of a high specific activity 35S ribosomal RNA precursor from Tetrahymena pyriformis and identification of its 5' terminus, pppAp. Biochemistry. 1978 Oct 31;17(22):4839–4844. doi: 10.1021/bi00615a035. [DOI] [PubMed] [Google Scholar]
  30. Niles E. G., Sutiphong J., Haque S. Structure of the Tetrahymena pyriformis rRNA gene. Nucleotide sequence of the transcription initiation region. J Biol Chem. 1981 Dec 25;256(24):12849–12856. [PubMed] [Google Scholar]
  31. Pelham H. R., Brown D. D. A specific transcription factor that can bind either the 5S RNA gene or 5S RNA. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4170–4174. doi: 10.1073/pnas.77.7.4170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Rosenberg M., Court D. Regulatory sequences involved in the promotion and termination of RNA transcription. Annu Rev Genet. 1979;13:319–353. doi: 10.1146/annurev.ge.13.120179.001535. [DOI] [PubMed] [Google Scholar]
  33. Rothblum L. I., Reddy R., Cassidy B. Transcription initiation site of rat ribosomal DNA. Nucleic Acids Res. 1982 Nov 25;10(22):7345–7362. doi: 10.1093/nar/10.22.7345. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rubtsov P. M., Musakhanov M. M., Zakharyev V. M., Krayev A. S., Skryabin K. G., Bayev A. A. The structure of the yeast ribosomal RNA genes. I. The complete nucleotide sequence of the 18S ribosomal RNA gene from Saccharomyces cerevisiae. Nucleic Acids Res. 1980 Dec 11;8(23):5779–5794. doi: 10.1093/nar/8.23.5779. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Saiga H., Mizumoto K., Matsui T., Higashinakagawa T. Determination of the transcription initiation site of Tetrahymena pyriformis rDNA using in vitro capping of 35S pre-rRNA. Nucleic Acids Res. 1982 Jul 24;10(14):4223–4236. doi: 10.1093/nar/10.14.4223. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sakonju S., Bogenhagen D. F., Brown D. D. A control region in the center of the 5S RNA gene directs specific initiation of transcription: I. The 5' border of the region. Cell. 1980 Jan;19(1):13–25. doi: 10.1016/0092-8674(80)90384-0. [DOI] [PubMed] [Google Scholar]
  37. Salim M., Maden B. E. Nucleotide sequence of Xenopus laevis 18S ribosomal RNA inferred from gene sequence. Nature. 1981 May 21;291(5812):205–208. doi: 10.1038/291205a0. [DOI] [PubMed] [Google Scholar]
  38. Sollner-Webb B., Reeder R. H. The nucleotide sequence of the initiation and termination sites for ribosomal RNA transcription in X. laevis. Cell. 1979 Oct;18(2):485–499. doi: 10.1016/0092-8674(79)90066-7. [DOI] [PubMed] [Google Scholar]
  39. Tsai S. Y., Tsai M. J., O'Malley B. W. Specific 5' flanking sequences are required for faithful initiation of in vitro transcription of the ovalbumin gene. Proc Natl Acad Sci U S A. 1981 Feb;78(2):879–883. doi: 10.1073/pnas.78.2.879. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Urano Y., Kominami R., Mishima Y., Muramatsu M. The nucleotide sequence of the putative transcription initiation site of a cloned ribosomal RNA gene of the mouse. Nucleic Acids Res. 1980 Dec 20;8(24):6043–6058. doi: 10.1093/nar/8.24.6043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  41. Veldman G. M., Klootwijk J., de Regt V. C., Planta R. J., Branlant C., Krol A., Ebel J. P. The primary and secondary structure of yeast 26S rRNA. Nucleic Acids Res. 1981 Dec 21;9(24):6935–6952. doi: 10.1093/nar/9.24.6935. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Verbeet M. P., Klootwijk J., van Heerikhuizen H., Fontijn R., Vreugdenhil E., Planta R. J. Molecular cloning of the rDNA of Saccharomyces rosei and comparison of its transcription initiation region with that of Saccharomyces carlsbergensis. Gene. 1983 Jul;23(1):53–63. doi: 10.1016/0378-1119(83)90216-0. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES