Functional Analysis of the Upstream Sequence of PHO81 Gene of Saccharomyces cerevisiae.

ABSTRACT

Deletion analysis on the fused PHO81-lacZ gene revealed two important regions in the upstream sequences of PHO81 gene -401 -289 bp and -1 012 -801 bp. They did not share higher similarity with the upstream regions of PHO5 and PHO84 gene, except that -401 -289 bp contains the 5'-CACGTG/T-3' motif, which was found among the upstream regions of PHO5 and PHO84 gene and was the core sequence of PHO4 binding site it also contains A/T-rich segments flanking the motif, which may be PHO2 binding sites. This suggests that the -401 -289 bp of the upstream region of PHO81 gene may contain upstream activation sequence (UAS), and -1 012 -801 bp may contain upstream enhancer sequence. The gel retardation assays of the -1 012 -801 bp was performed using yeast total protein extract, and the results showed that there was an unknown protein factor binding at the region.