Datin, a yeast poly(dA:dT)-binding protein, behaves as an activator of the wild-type ILV1 promoter and interacts synergistically with Reb1p.
A cis-acting element required for GCN4-independent basal-level transcription of ILV1 was previously identified in our laboratories as a binding site for the REB1 protein (Reb1p). Further deletion analysis of the ILV1 promoter region identified a second element also required for GCN4-independent basal-level ILV1 expression. This second element is an A.T-rich tract (26 As out of 32 nucleotides) situated 15 bp downstream of the Reb1p-binding site. Deletion of both the Reblp site and the poly(dA:dT) element totally eliminates basal activity of the ILV1 promoter. We show that the two elements act synergistically to control ILV1 expression and that the synergistic effect is distance dependent. We demonstrate that (i) datin (Dat1p), the only known poly (dA:dT)-binding protein in yeast, specifically binds to the ILV1 poly(dA:dT) element in vitro; (ii) Dat1p functions as a trans-activating factor in the ILV1 context; and (iii) the synergistic activation observed in vivo between the Reb1p site and the poly(dA:dT) element depends on the presence of the structural gene for Dat1p, DAT1.
Moreira
J M
JM
Department of Genetics, Institute of Molecular Biology, University of Copenhagen, Denmark.
Remacle
J E
JE
Kielland-Brandt
M C
MC
Holmberg
S
S
D001483:Base Sequence; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000502:physiology*; D050483:Dopamine Plasma Membrane Transport Proteins; D005656:Fungal Proteins / Q000235:genetics*; D005786:Gene Expression Regulation; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins; D011067:Poly dA-dT / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D029701:Saccharomyces cerevisiae Proteins; D013913:Threonine Dehydratase / Q000235:genetics*
C060858:DAT1 protein, S cerevisiae; D004268:DNA-Binding Proteins; D050483:Dopamine Plasma Membrane Transport Proteins; D005656:Fungal Proteins; D008562:Membrane Glycoproteins; D026901:Membrane Transport Proteins; D009419:Nerve Tissue Proteins; D029701:Saccharomyces cerevisiae Proteins; D011067:Poly dA-dT; D013913:Threonine Dehydratase