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Nucleic Acids Research, 1988, Vol. 16, No. 14 6693-6711
© 1988

Articles

Sequence-directed bends of DNA helix axis at the upstream activation sites of {alpha}-cell-specific genes in yeast

Kaoru Inokuchi*, Akiko Nakayama and Fumio Hishinuma

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

*To whom correspondence should be addressed

Received May 24, 1988. Accepted June 20, 1988.

The MF{alpha}1 gene of Saccharomyces cerevisiae, a major structural gene for mating pheromone {alpha} factor, is an {alpha}-cell-specific gene whose expression is regulated by the mating-type locus, MAT. Two upstream activation sites (UASMF{alpha}1s), which are binding sites for an activator protein, MAT{alpha}1 , mediate {alpha}-cell-specific expression of this gene. We show here that DNA fragments containing the UASMF{alpha}1 region exhibited anomalous slow electrophoretic mobilities on gels at lower temperature, but not at higher temperature, that is characteristic of bent DNA. We confirmed the sequence-directed bend at the UASMF{alpha}1 region by employing a circular permutation analysis and a DNA cyclization assay. Deletion analyses revealed the existence of two bends in this region, each of which overlaps each UASMF{alpha}1 element. The two bends were almost in phase; they lie in a nearly same plane with a same direction. We also show the existence of sequence-directed bend at the UAS region of the STE3 gene, another {alpha}-specific gene in S. cerevisiae. These bent DNAs may be involved in transcriptional regulation of this set of genes.


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