The Swi5 zinc-finger and Grf10 homeodomain proteins bind DNA cooperatively at the yeast HO promoter

Proc Natl Acad Sci U S A. 1993 Dec 1;90(23):11237-41. doi: 10.1073/pnas.90.23.11237.

Abstract

SWI5 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.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Amino Acid Sequence
  • Cell Cycle Proteins*
  • DNA-Binding Proteins / chemistry*
  • Deoxyribonucleoproteins / chemistry
  • Fungal Proteins / metabolism*
  • Gene Expression Regulation, Fungal*
  • Genes, Fungal*
  • Genes, Homeobox
  • Homeodomain Proteins*
  • Macromolecular Substances
  • Molecular Sequence Data
  • Promoter Regions, Genetic
  • Saccharomyces cerevisiae / genetics*
  • Saccharomyces cerevisiae Proteins*
  • Trans-Activators / metabolism*
  • Transcription Factors / metabolism*
  • Zinc Fingers

Substances

  • Cell Cycle Proteins
  • DNA-Binding Proteins
  • Deoxyribonucleoproteins
  • Fungal Proteins
  • Homeodomain Proteins
  • Macromolecular Substances
  • PHO2 protein, S cerevisiae
  • SWI5 protein, S cerevisiae
  • Saccharomyces cerevisiae Proteins
  • Trans-Activators
  • Transcription Factors