Nucleic Acids Research, Vol 24, Issue 22 4479-4486, Copyright © 1996 by Oxford University Press
S Barbaric, M Munsterkotter, J Svaren and W Horz
Two transcription factors, the bHLH protein Pho4 and the homeodomain
protein Pho2, are required for transcriptional activation of the PHO5
promoter in Saccharomyces cerevisiae. There are two essential Pho4 binding
sites, corresponding to the regulatory elements UASp1 and UASp2 at the PHO5
promoter, but only a single, dispensable Pho2 binding site had previously
been identified. We have reinvestigated binding of Pho2 to the PHO5
promoter using purified recombinant protein and have found multiple Pho2
binding sites of different affinities along the promoter. One of the high
affinity Pho2 sites largely overlaps the Pho4 binding site at UASp1.
Cooperative DNA binding of the two proteins to their overlapping sites,
resulting in a high-affinity ternary complex, was demonstrated. Pho2 and
Pho4 also bind DNA cooperatively at UASp2 where two Pho2 sites flank the
Pho4 site. Finally, Pho2 facilitates binding of Pho4 to a third, cryptic
Pho4 binding site which binds Pho4 with lower affinity than UASp1 or UASp2.
These results suggest that cooperative DNA binding with Pho4 is integral to
the mechanism by which Pho2 regulates transcription of the PHO5 gene.
ARTICLES
The homeodomain protein Pho2 and the basic-helix-loop-helix protein Pho4 bind DNA cooperatively at the yeast PHO5 promoter
Institut fur Physiologische Chemie, Universitat Munchen, Germany.
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