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Isolation of Drosophila proteins that bind selectively to left-handed Z-DNA
Journal article   Open access   Peer reviewed

Isolation of Drosophila proteins that bind selectively to left-handed Z-DNA

Alfred Nordheim, Paul Tesser, Fernando Azorin, Young Ha Kwon, Achim Möller and Alexander Rich
Proceedings of the National Academy of Sciences of the United States of America, Vol.79(24 I), pp.7729-7733
1982
DOI: 10.1073/pnas.79.24.7729
PMID: 6296848
url
https://doi.org/10.1073/pnas.79.24.7729View
Published (Version of record) Open Access

Abstract

An affinity column for isolating Z-DNA binding proteins was made by attaching brominated poly(dG-dC) to Sephadex. Proteins from Drosophila nuclei were prepared and those that could bind to Escherichia coli B-DNA were removed from the solution. The remaining proteins were passed over the Z-DNA affinity column and then eluted with NaCl. Using both direct and competitive filter binding assays, we found that the eluted proteins bind to brominated poly(dG-dC) (Z-DNA) and poly(dG-m5dC) but not to poly(dG-dC) (B-DNA), native or denatured E. coli or calf thymus DNA, or brominated oligonucleotides. The proteins also bind to negatively supercoiled plasmids carrying Z-DNA sequences but not to relaxed or linearized plasmids in which the Z-DNA conformation is no longer present. Gel analysis reveals a mixture of several large proteins up to approximately 150,000 daltons.

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