Z-DNA BINDING-PROTEIN FROM CHICKEN BLOOD NUCLEI

A protein (Zalpha) that appears to be highly specific for the left-han ded Z-DNA conformer has been identified in chicken blood nuclear extra cts. Zalpha activity is measured in a band-shift assay by using a radi oactive probe consisting of a (dC-dG)35 oligomer that has 50% of the d eoxycytosines replaced with 5-bromodeoxycytosine. In the presence of 1 0 mM Mg2+, the probe converts to the Z-DNA conformation and is bound b y Zalpha. The binding of Zalpha to the radioactive probe is specifical ly blocked by competition with linear poly(dC-dG) stabilized in the Z- DNA form by chemical bromination but not by B-form poly(dC-dG) or boil ed salmon-sperm DNA. In addition, the binding activity of Zalpha is co mpetitively blocked by supercoiled plasmids containing a Z-DNA insert but not by either the linearized plasmid or by an equivalent amount of the parental supercoiled plasmid without the Z-DNA-forming insert. Za lpha can be crosslinked to the P-32-labeled brominated probe with UV l ight, allowing us to estimate that the minimal molecular mass of Zalph a is 39 kDa.