DNA binding properties of theSaccharomyces cerevisiae DAT1gene product
| Autor: | Donna Gordon, Edward Winter, Brian J. Reardon, Michelle J. Ballard |
|---|---|
| Rok vydání: | 1995 |
| Předmět: |
Dopamine Plasma Membrane Transport Proteins
Membrane Glycoproteins Saccharomyces cerevisiae Proteins HMG-box Oligonucleotide Binding protein Genetic Vectors Membrane Transport Proteins bZIP domain Nerve Tissue Proteins DNA-binding domain Biology Binding Competitive DNA-Binding Proteins Fungal Proteins DNA binding site Biochemistry Escherichia coli Genetics Binding site DNA Probes DNA Fungal Binding domain |
| Zdroj: | Nucleic Acids Research. 23:4900-4906 |
| ISSN: | 1362-4962 0305-1048 |
| DOI: | 10.1093/nar/23.23.4900 |
| Popis: | The DAT1 gene of Saccharomyces cerevisiae encodes a DNA binding protein (Dat1p) that specifically recognizes the minor groove of non-alternating oligo(A).oligo(T) tracts. Sequence-specific recognition requires arginine residues found within three perfectly repeated pentads (G-R-K-P-G) of the Dat1p DNA binding domain [Reardon, B. J., Winters, R. S., Gordon, D., and Winter, E. (1993) Proc. Natl. Acad. Sci. USA 90, 11327-1131]. This report describes a rapid and simple method for purifying the Dat1p DNA binding domain and the biochemical characterization of its interaction with oligo(A).oligo(T) tracts. Oligonucleotide binding experiments and the characterization of yeast genomic Dat1p binding sites show that Dat1p specifically binds to any 11 base sequence in which 10 bases conform to an oligo(A).oligo(T) tract. Binding studies of different sized Dat1p derivatives show that the Dat1p DNA binding domain can function as a monomer. Competition DNA binding assays using poly(I).poly(C), demonstrate that the minor groove oligo(A).oligo(T) constituents are not sufficient for high specificity DNA binding. These data constrain the possible models for Dat1p/oligo(A).oligo(T) complexes, suggest that the DNA binding domain is in an extended structure when complexed to its cognate DNA, and show that Dat1p binding sites are more prevalent than previously thought. |
| Databáze: | OpenAIRE |
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