Nuclease activity of 1,10-phenanthroline-copper ion. Conformational analysis and footprinting of the lac operon.
Nuclease activity of 1,10-phenanthroline-copper ion. Conformational analysis and footprinting of the lac operon.
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1,10-菲咯啉-铜离子的核酸酶活性。
DOI：
10.1021/bi00348a032
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发表时间：
1985
期刊：
影响因子：
2.9
通讯作者：
D. Sigman
中科院分区：
文献类型：
--
作者：
A. Spassky;D. Sigman
1，10-菲咯啉-铜[（OP）2Cu+]的核酸酶活性优先切割保守启动子特异性序列（Pribnow框）中的野生型、Ps和L 8-UV-5 lac启动子。该区域内野生型片段的优选攻击位点在模板链上的位置-13和-12。当用（OP）2Cu+切割来自Ps启动子的可比较片段（其在位置-9处不同于野生型（T而不是C））时，凝胶模式中位置-10处的新的强条带是清楚的。还可以观察到位置-11处的切割明显增加。Ps启动子向L 8-UV-5启动子的转化（位置-8处从A变为T，位置-66处从C变为T）导致位置-13至-10处的四个显著条带的相对强度改变。最值得注意的是，位置-10处的强度在L 8-UV-5中衰减。如果分析错义链的切割，则Pribnow盒区域对配位复合物的超敏性也是明显的。在这种情况下，强切口的区域范围为位置-11至-3，条带的相对强度取决于启动子的一级序列。这些数据表明，一个单一的碱基变化诱导DNA结构的局部变化。这种新的结构可能是启动子效率显著差异的原因。胰腺脱氧核糖核酸酶I（DNase I）相对于序列的其他区域不优先切割Pribnow盒。（250字处删节）
The nuclease activity of 1,10-phenanthroline-copper [(OP)2Cu+] preferentially nicks the wild-type, Ps, and L8-UV-5 lac promoters in the conserved promoter specific sequence (Pribnow box). The preferred sites of attack of the wild-type fragment within this region are at positions -13 and -12 on the template strand. When the comparable fragment from the Ps promoter, which differs from the wild type at position -9 (T instead of C), is cleaved with (OP)2Cu+, a new strong band at position -10 in the gel patterns is clear. An apparent increase in cutting at position -11 can also be observed. The conversion of the Ps promoter to the L8-UV-5 promoter (a change from an A to a T at position -8 and a change from a C to a T at position -66) results in alteration of the relative intensities of the four prominent bands at positions -13 to -10. Most notably, the intensity at position -10 is attenuated in L8-UV-5. The hypersensitivity of the Pribnow box region to the coordination complex is also apparent if the cutting of the missense strand is analyzed. The region of strong nicking in this case ranges from positions -11 to -3, and the relative intensities of the bands depend on the primary sequence of the promoters. These data suggest that a single base change induces local variation in the DNA structure. This new structure may be responsible for the notable difference in the efficiency of the promoters. Pancreatic deoxyribonuclease I (DNase I) does not preferentially cleave the Pribnow box relative to other regions of the sequence.(ABSTRACT TRUNCATED AT 250 WORDS)