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Efficient expression and Zn(II)-dependent structure of the DNA binding domain of the yeast GAL4 protein
Y Serikawa1, M Shirakawa, H Matsuo
1Institute for Protein Research, Osaka University, Japan.
Abstract:
Three protein fragments of different sizes which contain the DNA binding domain of transcription factor GAL4 from Saccharomyces cerevisiae have been expressed in functional forms in Escherichia coli. DNase I footprinting and gel retardation assays showed that the purified proteins bound to the same DNA sequence on the gal1-gal10 promoter as intact GAL4 does. Denaturation--refolding experiments demonstrated that Zn(II) is necessary for maintenance of the conformation of the DNA binding domain of GAL4, as judged on UV-CD and 1H-NMR measurements, as well as for specific DNA binding.
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