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The Swi5 Zinc-Finger and Grf10 Homeodomain Proteins Bind DNA Cooperatively at the Yeast HO Promoter
Robert M. Brazas and David J. Stillman
Proceedings of the National Academy of Sciences of the United States of America
Vol. 90, No. 23 (Dec. 1, 1993), pp. 11237-11241
Published by: National Academy of Sciences
Stable URL: http://www.jstor.org/stable/2363684
Page Count: 5
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SW15 encodes a zinc-finger protein required for expression of the yeast HO gene. Using Swi5 protein that was purified from a bacterial expression system, we previously isolated a yeast factor that stimulates binding of Swi5 to the HO promoter. N-terminal amino acid sequence analysis identified the Swi5 stimulatory factor as the product of the GRF10 gene, which encodes a yeast homeodomain protein. GRF10, also known as PHO2 and BAS2, is a transcriptional activator of the PHO5 acid phosphatase gene and the HIS4 histidine biosynthesis gene. Grf10 protein purified from a bacterial expression system binds DNA cooperatively with Swi5 in vitro. Analysis of disassociation rates indicates that the Grf10-Swi5-DNA complex has a longer half-life than protein-DNA complexes that contain only Swi5 or Grf10. Finally, we show that HO expression is reduced in yeast strains containing grf10 null mutations and that full expression of a heterologous promoter containing a SWI5-dependent HO upstream activation sequence element requires GRF10.
Proceedings of the National Academy of Sciences of the United States of America © 1993 National Academy of Sciences