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

S Firek, C Read, D R Smith, T Moss
Author Information
  1. S Firek: Biophysics Laboratories, Portsmouth Polytechnic, UK.

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.

References

  1. Nucleic Acids Res. 1985 Sep 11;13(17):6237-48 [PMID: 2995922]
  2. Nucleic Acids Res. 1982 Nov 25;10(22):7345-62 [PMID: 6296773]
  3. Mol Cell Biol. 1986 Jan;6(1):227-35 [PMID: 3785147]
  4. Nucleic Acids Res. 1987 Sep 25;15(18):7429-41 [PMID: 3658698]
  5. Proc Natl Acad Sci U S A. 1988 Feb;85(3):669-73 [PMID: 3422449]
  6. Science. 1988 Sep 2;241(4870):1192-7 [PMID: 3413483]
  7. Nucleic Acids Res. 1988 Nov 25;16(22):10657-68 [PMID: 3205719]
  8. Oxf Surv Eukaryot Genes. 1985;2:207-50 [PMID: 3916926]
  9. Mol Cell Biol. 1989 Sep;9(9):3777-84 [PMID: 2779566]
  10. Cell. 1977 Nov;12(3):721-32 [PMID: 922889]
  11. Proc Natl Acad Sci U S A. 1978 May;75(5):2170-4 [PMID: 209457]
  12. Cell. 1982 Oct;30(3):835-42 [PMID: 7139716]
  13. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6908-11 [PMID: 6294665]
  14. Nucleic Acids Res. 1985 May 24;13(10):3515-32 [PMID: 2989774]
  15. Mol Cell Biol. 1985 Mar;5(3):554-62 [PMID: 3990683]
  16. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2137-41 [PMID: 6326106]
  17. Proc Natl Acad Sci U S A. 1984 Jan;81(2):299-303 [PMID: 6320178]
  18. Cell. 1983 Nov;35(1):199-206 [PMID: 6684995]
  19. Proc Natl Acad Sci U S A. 1983 Jun;80(12):3558-62 [PMID: 6304717]
  20. Proc Natl Acad Sci U S A. 1983 Jun;80(11):3265-8 [PMID: 6407009]
  21. Nucleic Acids Res. 1983 May 25;11(10):3317-32 [PMID: 6304628]
  22. Proc Natl Acad Sci U S A. 1986 Feb;83(3):604-8 [PMID: 3456157]

MeSH Term

Animals
Base Sequence
Cloning, Molecular
DNA Mutational Analysis
DNA, Ribosomal
Molecular Sequence Data
Promoter Regions, Genetic
RNA Polymerase I
RNA, Ribosomal
Sequence Homology, Nucleic Acid
Substrate Specificity
Xenopus laevis

Chemicals

DNA, Ribosomal
RNA, Ribosomal
RNA Polymerase I

Word Cloud

Created with Highcharts 10.0.0promotercoreregionXenopuslaevisRNApointmutationanalysistranscriptionboundary+4-33suggestparticularnucleotides-7interactionspre-ribosomalsubjectedtotal27mutantswithin78basepair-64+14relativestart+1assayedoocytemicroinjectionresultslocate3'5'-39generallysimilarorganisationmammalianpromotersconservedguanidine-16clearlyessentialfunctiondatamachineryoccurfourdistinctregionsaround+2-17-20-28periodicityfunctionallyimportantconsistentmodelprotein-DNAtakeplacepredominantlyonesideDNAhelixPointpolymerase

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