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Nucleic Acids Research, 1988, Vol. 16, No. 4 1407-1422
© 1988

Articles

The protein factor which binds to the upstream activating sequence of Saccharomyces cerevisiae ENO1 gene

Masayuki Machida*, Hiroshi Uemura, Yoshifumi Jigami and Hideaki Tanaka

National Chemical Laboratory for Industry, Kagaku Gijutsu Kenkyusho, Tsukuba Research Center Yatabe, Ibaraki 305, Japan

*To whom correspondence should be addressed

Received October 7, 1987. Revised January 20, 1988. Accepted January 20, 1988.

Using a gel retardation assay it was shown that the 87 bp DNA fragment (UAS87) containing the upstream activating sequence (UAS) of S. cerevisiae ENO1 gene and a nuclear extract gave rise to three migration-retarded species apecific to UAS87. Heat- or proteinase-treatment of the nuclear extract revealed that these species were protein-DNA complexes. The precise binding region of the protein identified by DNaseI protection analysis was found to include a CCAAACA sequence which forms a dyad-symmetrical structure. The amount of one of the three migration-retarded species significantly increased when cells were grown in medium containing a gluconeogenic carbon source. The introduction of pGCR8, a multicopy plasmid containing GCR1 gene, a regulatory gene controling the expression of several glycolytic enzymes, showed no effect on the amount of three migration-retarded species.


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