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Nucleic Acids Research, 1989, Vol. 17, No. 18 7427-7439
© 1989

MOLECULAR BIOLOGY

The extended promoter of the gene encoding ribosomal protein S33 in yeast consists of multiple protein binding elements

Martein H. Herruer, Willem H. Mager, Thijs M. Doorenbosch, Peter L. M. Wessels, Trudy M. Wassenaar and Rudi J Planta*

Biochemisch Laboratorium, Vrije Universiteit de Boelelaan 1083, 1081 HV Amsterdam, The Netherlands

*To whom correspondence should be addressed

Received July 6, 1989. Revised August 11, 1989. Accepted August 11, 1989.

At least 4 different, protein binding cis-acting elements are present in the upstream region of the S33-gene. The major protein binding site is situated between positions –148 and –163 relative to the ATG start codon. It binds a trans-acting factor designated SUF (S33 Upstream Factor). When yeast cells are growing on glucose, deletion of this site results in a decrease of transcription of 50%. Using ethanol as a carbon-source, deletion of the SUF-responsive site lowers the transcription as much as 80%. A second protein binding site is found between positions –85 and –105. Only extracts from glucose-grown cells contain a factor that is able to bind to this site in vitro. A third protein binding site was found using a protein extract from ethanol-grown cells. This site, which is located quite close to the transcriptional start site, is probably responsible for the 20% residual transcription when the SUF binding site is removed. Finally, a site far upstream was found, which binds a protein from both glucose-grown and ethanol-grown cells. This site may function as an upstream repression site which is only functional when a non-fermentable carbon-source is used. Taking these findings into account, we present a model for the carbon-source dependent transcription activation of the gene encoding S33.


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