Z-DNA BINDING-PROTEIN FROM CHICKEN BLOOD NUCLEI

被引:40
作者
HERBERT, AG [1 ]
SPITZNER, JR [1 ]
LOWENHAUPT, K [1 ]
RICH, A [1 ]
机构
[1] MIT,DEPT BIOL,77 MASSACHUSETTS AVE,CAMBRIDGE,MA 02139
关键词
BAND-SHIFT; UV CROSS-LINK; SUPERCOILED DNA;
D O I
10.1073/pnas.90.8.3339
中图分类号
O [数理科学和化学]; P [天文学、地球科学]; Q [生物科学]; N [自然科学总论];
学科分类号
07 ; 0710 ; 09 ;
摘要
A protein (Zalpha) that appears to be highly specific for the left-handed Z-DNA conformer has been identified in chicken blood nuclear extracts. Zalpha activity is measured in a band-shift assay by using a radioactive probe consisting of a (dC-dG)35 oligomer that has 50% of the deoxycytosines replaced with 5-bromodeoxycytosine. In the presence of 10 mM Mg2+, the probe converts to the Z-DNA conformation and is bound by Zalpha. The binding of Zalpha to the radioactive probe is specifically 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 boiled salmon-sperm DNA. In addition, the binding activity of Zalpha is competitively 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. Zalpha can be crosslinked to the P-32-labeled brominated probe with UV light, allowing us to estimate that the minimal molecular mass of Zalpha is 39 kDa.
引用
收藏
页码:3339 / 3342
页数:4
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