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. 1994 Apr 25;22(8):1450-5.
doi: 10.1093/nar/22.8.1450.

A simple in vivo footprinting method to examine DNA-protein interactions over the yeast PYK UAS element

I Dumitru  1 , J B McNeil
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Free PMC article

A simple in vivo footprinting method to examine DNA-protein interactions over the yeast PYK UAS element

I Dumitru et al. Nucleic Acids Res. .
Free PMC article

Abstract

In this report a modification to the in vivo footprinting assay is described. The method includes dimethyl sulfate treatment of whole yeast cells, followed by reiterative primer extension of the methylated genomic DNA using Taq DNA polymerase. Under appropriate reaction conditions chain extension terminates opposite a methylated purine when Taq DNA polymerase encounters a modified adenine or guanine. The procedure was used to examine, in vivo DNA-protein contacts over the upstream activation site (UAS) of the Saccharomyces cerevisiae PYK gene. In vivo analysis, using isogenic strains of yeast and Escherichia coli transformed with plasmid DNAs, confirmed the binding of both the trans-acting factor RAP1 and the transcriptional activator GCR1 to cis-acting recognition sites located within the PYK UAS element.

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References

    1. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463-7 - PubMed
    1. Biochem J. 1963 Oct;89:127-38 - PubMed
    1. Methods Enzymol. 1980;65(1):499-560 - PubMed
    1. J Biol Chem. 1981 Dec 25;256(24):13074-8 - PubMed
    1. Proc Natl Acad Sci U S A. 1984 Apr;81(7):1991-5 - PubMed

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