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Nucleic Acids Research, 2001, Vol. 29, No. 13 e64
© 2001 Oxford University Press

The mapping of nucleosomes and regulatory protein binding sites at the Saccharomyces cerevisiae MFA2 gene: a high resolution approach

Yumin Teng, Shirong Yu and Raymond Waters* School of Biological Sciences, University of Wales Swansea, Singleton Park, Swansea SA2 8PP, UK

We have developed an end-labelling approach to map the positions of nucleosomes and protein binding sites at nucleotide resolution by footprinting micrococcal nuclease (MNase)-sensitive sites. Using this approach we determined that the MFA2 gene and its upstream control regions have four positioned nucleosomes when transcription is repressed in mating type {alpha} cells and that the nucleosomes lose their positioning when the gene became transcriptionally active in mating type a cells. We also detected MNase-hypersensitive sites in the {alpha}2 operator region of MFA2 in {alpha} cells but not in a cells. These probably result from the change in the local DNA conformation due to protein(s) binding in this region that governs MFA2 transcription.

* To whom correspondence should be addressed. Tel: +44 1792 295384; Fax: +44 1792 295447; Email: r.waters{at}swansea.ac.uk


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