Isolation and Characterization of Z-DNA Binding Proteins from Wheat Germ

Eileen M. Lafer, Rui Sousa, Barry Rosen, Anna Hsu, Alexander Rich

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

The preparation of a heterogeneous non-histone protein extract from wheat germ utilizing Br-poly(dG-dC)-poly(dG-dC) (Z-DNA) affinity chromatography is described. The binding characteristics of antibodies against Z-DNA are used as a model system to define important criteria that the DNA binding behavior of a Z-DNA binding protein should display. We show that the wheat germ extract contains DNA binding proteins specific for left-handed Z-DNA by these criteria. The affinity of the proteins measured by competition experiments was approximately 105 greater for Br-poly(dG-dC)-poly(dG-dC) (Z-DNA) than for poly(dG-dC)-poly(dG-dC) (B-DNA). The affinity of the proteins for plasmid DNA increases with increasing negative superhelicity which is known to stabilize Z-DNA. The proteins are shown to compete with Z-DNA antibodies for binding to supercoiled plasmids. Finally, the affinity for two plasmids at a given superhelical density is greater for the plasmid containing an insert known to form Z-DNA than for a plasmid without the insert. The proteins exhibit a 2-3-fold greater affinity for stretches of (dC-dA)n-(dT-dG)n over stretches of (dG-dC)n-(dG-dC)n when both sequences are induced to form Z-DNA by supercoiling.

Original languageEnglish (US)
Pages (from-to)5070-5076
Number of pages7
JournalBiochemistry
Volume24
Issue number19
DOIs
StatePublished - Sep 1 1985

ASJC Scopus subject areas

  • Biochemistry

Fingerprint Dive into the research topics of 'Isolation and Characterization of Z-DNA Binding Proteins from Wheat Germ'. Together they form a unique fingerprint.

  • Cite this

    Lafer, E. M., Sousa, R., Rosen, B., Hsu, A., & Rich, A. (1985). Isolation and Characterization of Z-DNA Binding Proteins from Wheat Germ. Biochemistry, 24(19), 5070-5076. https://doi.org/10.1021/bi00340a017