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Nucleic Acids Research, 2002, Vol. 30, No. 19 4199-4207
© 2002 Oxford University Press

Transcription initiation in vivo without classical transactivators: DNA kinks flanking the core promoter of the housekeeping yeast adenylate kinase gene, AKY2, position nucleosomes and constitutively activate transcription

Michaela Angermayr*, Ulrich Oechsner, Kerstin Gregor, Gary P. Schroth and Wolfhard Bandlow

Department Biologie I, Bereich Genetik, Ludwig-Maximilians-Universität München, Maria-Ward-Strasse 1a, D-80638 Munich, Germany

*To whom correspondence should be addressed. Tel: +49 89 2180 6176; Fax: +49 89 2180 6160; Email: m.angermayr{at}lrz.uni-muenchen.de
Present addresses:
Ulrich Oechsner, Lichtenstein Pharmaceutica GmbH und Co., Industriestrasse 10, D-82256 Fürstenfeldbruck, Germany
Gary P. Schroth, Genelabs Technologies Inc., 505 Penobscot Drive, Redwood City, CA 94063, USA

The housekeeping gene of the major adenylate kinase in Saccharomyces cerevisiae (AKY2, ADK1) is constitutively transcribed at a moderate level. The promoter has been dissected in order to define elements that effect constitutive transcription. Initiation of mRNA synthesis at the AKY2 promoter is shown to be mediated by a non-canonic core promoter, (TA)6. Nucleotide sequences 5' of this element only marginally affect transcription suggesting that promoter activation can dispense with transactivators and essentially involves basal transcription. We show that the core promoter of AKY2 is constitutively kept free of nucleosomes. Analyses of permutated AKY2 promoter DNA revealed the presence of bent DNA. DNA structure analysis by computer and by mutation identified two kinks flanking an interstitial stretch of 65 bp of moderately bent core promoter DNA. Kinked DNA is likely incompatible with packaging into nucleosomes and responsible for positioning nucleosomes at the flanks allowing unimpeded access of the basal transcription machinery to the core promoter. The data show that in yeast, constitutive gene expression can dispense with classical transcriptional activator proteins, if two prerequisites are met: (i) the core promoter is kept free of nucleosomes; this can be due to structural properties of the DNA as an alternative to chromatin remodeling factors; and (ii) the core promoter is pre-bent to allow a high rate of basal transcription initiation.


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