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Nucleic Acids Research, 1991, Vol. 19, No. 11 3099-3103
© 1991

MOLECULAR BIOLOGY

Lack of a requirement for strict rotational alignment among transcription factor binding sites in yeast

Kaoru Inokuchi* and Akiko Nakayama

Laboratory of Molecular Genetics, Mitsubishi Kasei Institute of Life Sciences 11 Minamiooya, Machida-shi, Tokyo 194, Japan

*To whom correspondence should be addressed at Center for Neurobiology and Behavior, College of Physicians and Surgeons, Columbia University, 722 West 168th Street, New York, NY 10032, USA

Received December 28, 1990. Revised April 23, 1991. Accepted April 23, 1991.

The MF{alpha}1 gene of Saccharomyces cerevisiae is an {alpha} specific gene whose expression is regulated by two upstream activation sites (UASMF{alpha}1s), which are the binding sites for activator proteins, MAT{alpha}1 and MCM1. UASMF{alpha}1 elements act in a synergistic fashion; lack of either element causes reduced expression levels that are 6- to 45-fold less than that of intact promoter in {alpha} cells. We investigated the effect of manipulating rotational alignments among transcription factor binding sites of MF{alpha}1 on the expression of the gene. The expression level of MF{alpha}1 decreased with the spacing between the two UASMF{alpha}1s and no synergism was observed when the distance of the two elements was longer than 90 base pairs. No strict rotational alignment was required for their synergistic action. We also show that the spacing between UASMF{alpha}1 elements and TFIID binding site, the TATA box, had little, if any, effect on MF{alpha}1 expression. We chose a sufficient number of positions for UASMF{alpha}1s to ensure that, in several of these positions, MAT{alpha}1 and MCM1 were on the opposite side of the DNA helix with respect to the TATA box.


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