Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1990 Jan 11;18(1):105–109. doi: 10.1093/nar/18.1.105

Point mutation analysis of the Xenopus laevis RNA polymerase I core promoter.

S Firek 1, C Read 1, D R Smith 1, T Moss 1
PMCID: PMC330209  PMID: 2308816

Abstract

The core region of the Xenopus laevis pre-ribosomal RNA promoter was subjected to point mutation analysis. A total of 27 point mutants within a 78 base pair region from -64 to +14, (relative to the start of transcription at +1), were assayed by oocyte microinjection. The results locate the 3' boundary of the core promoter at +4 and the 5' boundary at between -33 and -39 and suggest that this region of the Xenopus promoter is generally similar in organisation to mammalian core promoters. In particular, the conserved guanidine nucleotides at -7 and -16 are clearly essential for promoter function. The data suggest that interactions between the transcription machinery and the promoter occur in four distinct regions around +2 to +4, -7, -17 to -20 and -28 to -33. This particular periodicity of functionally important nucleotides is consistent with a model in which all protein-DNA interactions take place from predominantly one side of the DNA helix.

Full text

PDF
105

Images in this article

106Image
on p.106

Selected References

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

  1. Bell S. P., Learned R. M., Jantzen H. M., Tjian R. Functional cooperativity between transcription factors UBF1 and SL1 mediates human ribosomal RNA synthesis. Science. 1988 Sep 2;241(4870):1192–1197. doi: 10.1126/science.3413483. [DOI] [PubMed] [Google Scholar]
  2. Berk A. J., Sharp P. A. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease-digested hybrids. Cell. 1977 Nov;12(3):721–732. doi: 10.1016/0092-8674(77)90272-0. [DOI] [PubMed] [Google Scholar]
  3. Clos J., Buttgereit D., Grummt I. A purified transcription factor (TIF-IB) binds to essential sequences of the mouse rDNA promoter. Proc Natl Acad Sci U S A. 1986 Feb;83(3):604–608. doi: 10.1073/pnas.83.3.604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Firek S., Read C., Smith D. R., Moss T. The Xenopus laevis ribosomal gene terminator contains sequences that both enhance and repress ribosomal transcription. Mol Cell Biol. 1989 Sep;9(9):3777–3784. doi: 10.1128/mcb.9.9.3777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Grummt I. Nucleotide sequence requirements for specific initiation of transcription by RNA polymerase I. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6908–6911. doi: 10.1073/pnas.79.22.6908. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Haltiner M. M., Smale S. T., Tjian R. Two distinct promoter elements in the human rRNA gene identified by linker scanning mutagenesis. Mol Cell Biol. 1986 Jan;6(1):227–235. doi: 10.1128/mcb.6.1.227. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Harrington C. A., Chikaraishi D. M. Identification and sequence of the initiation site for rat 45S ribosomal RNA synthesis. Nucleic Acids Res. 1983 May 25;11(10):3317–3332. doi: 10.1093/nar/11.10.3317. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jones M. H., Learned R. M., Tjian R. Analysis of clustered point mutations in the human ribosomal RNA gene promoter by transient expression in vivo. Proc Natl Acad Sci U S A. 1988 Feb;85(3):669–673. doi: 10.1073/pnas.85.3.669. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Kishimoto T., Nagamine M., Sasaki T., Takakusa N., Miwa T., Kominami R., Muramatsu M. Presence of a limited number of essential nucleotides in the promoter region of mouse ribosomal RNA gene. Nucleic Acids Res. 1985 May 24;13(10):3515–3532. doi: 10.1093/nar/13.10.3515. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kohorn B. D., Rae P. M. Localization of DNA sequences promoting RNA polymerase I activity in Drosophila. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3265–3268. doi: 10.1073/pnas.80.11.3265. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kownin P., Iida C. T., Brown-Shimer S., Paule M. R. The ribosomal RNA promoter of Acanthamoeba castellanii determined by transcription in a cell-free system. Nucleic Acids Res. 1985 Sep 11;13(17):6237–6248. doi: 10.1093/nar/13.17.6237. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Learned R. M., Smale S. T., Haltiner M. M., Tjian R. Regulation of human ribosomal RNA transcription. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3558–3562. doi: 10.1073/pnas.80.12.3558. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Miller K. G., Tower J., Sollner-Webb B. A complex control region of the mouse rRNA gene directs accurate initiation by RNA polymerase I. Mol Cell Biol. 1985 Mar;5(3):554–562. doi: 10.1128/mcb.5.3.554. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Moss T., Mitchelson K., de Winter R. The promotion of ribosomal transcription in eukaryotes. Oxf Surv Eukaryot Genes. 1985;2:207–250. [PubMed] [Google Scholar]
  15. Moss T. Transcription of cloned Xenopus laevis ribosomal DNA microinjected into Xenopus oocytes, and the identification of an RNA polymerase I promoter. Cell. 1982 Oct;30(3):835–842. doi: 10.1016/0092-8674(82)90288-4. [DOI] [PubMed] [Google Scholar]
  16. Reeder R. H., Pennock D., McStay B., Roan J., Tolentino E., Walker P. Linker scanner mutagenesis of the Xenopus laevis ribosomal gene promoter. Nucleic Acids Res. 1987 Sep 25;15(18):7429–7441. doi: 10.1093/nar/15.18.7429. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Shortle D., Nathans D. Local mutagenesis: a method for generating viral mutants with base substitutions in preselected regions of the viral genome. Proc Natl Acad Sci U S A. 1978 May;75(5):2170–2174. doi: 10.1073/pnas.75.5.2170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Skinner J. A., Ohrlein A., Grummt I. In vitro mutagenesis and transcriptional analysis of a mouse ribosomal promoter element. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2137–2141. doi: 10.1073/pnas.81.7.2137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Sollner-Webb B., Wilkinson J. A., Roan J., Reeder R. H. Nested control regions promote Xenopus ribosomal RNA synthesis by RNA polymerase I. Cell. 1983 Nov;35(1):199–206. doi: 10.1016/0092-8674(83)90222-2. [DOI] [PubMed] [Google Scholar]
  21. Walker P., Reeder R. H. The Xenopus laevis ribosomal gene promoter contains a binding site for nuclear factor-1. Nucleic Acids Res. 1988 Nov 25;16(22):10657–10668. doi: 10.1093/nar/16.22.10657. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Yamamoto O., Takakusa N., Mishima Y., Kominami R., Muramatsu M. Determination of the promoter region of mouse ribosomal RNA gene by an in vitro transcription system. Proc Natl Acad Sci U S A. 1984 Jan;81(2):299–303. doi: 10.1073/pnas.81.2.299. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES