Di Mauro E, Ballario P, Pedone F
Biochemistry. 1980 Apr 1;19(7):1392-6. doi: 10.1021/bi00548a020.
We describe the properties of the complexes that form between Escherichia coli RNA polymerase and Paracentrotus lividus DNA: dissociation kinetics, temperature dependence of the complex formation, resistance to heparin, and range of RNA polymerase-DNA weight/weight ratios that give rise to the stable binding events. The amount and distribution of the sites that form stable binding [class A sites as defined by Hinkle & Chamberlin [Hinkle, D., & Chamberlin, M. J. (1972) J. Mol. Biol. 70, 157]] with E. coli RNA polymerase were determined by the analysis of the dissociation of complexes formed by the enzyme on DNA fragments of various length. The P. lividus appears to form 3.1 X 10(5) stable (t1/2 greater than or equal to 15 min) complexes per haploid genome; the great majority of these complexes shows a short-range distribution (1000-2000 base pairs). The observed attributes of the stable binding sites of P. lividus DNA for E. coli RNA polymerase (amount, distribution, and quantitative ability to start in vitro RNA chains) point to the conclusion that E. coli and sea urchin DNA are nearly indistinguishable by the criteria adopted. The behavior of the sea urchin stable binding sites for the E. coli enzyme is not consistent with the expected behavior of the in vivo promoters.
我们描述了大肠杆菌RNA聚合酶与紫海胆DNA之间形成的复合物的特性:解离动力学、复合物形成的温度依赖性、对肝素的抗性以及导致稳定结合事件的RNA聚合酶与DNA重量/重量比范围。通过分析该酶与各种长度DNA片段形成的复合物的解离,确定了与大肠杆菌RNA聚合酶形成稳定结合的位点(如Hinkle和Chamberlin所定义的A类位点[Hinkle, D., & Chamberlin, M. J. (1972) J. Mol. Biol. 70, 157])的数量和分布。紫海胆每个单倍体基因组似乎形成3.1×10⁵个稳定(半衰期大于或等于15分钟)的复合物;这些复合物中的绝大多数显示出短程分布(1000 - 2000个碱基对)。紫海胆DNA对大肠杆菌RNA聚合酶稳定结合位点的观察特性(数量、分布以及体外启动RNA链的定量能力)表明,按照所采用的标准,大肠杆菌和海胆DNA几乎无法区分。海胆DNA对大肠杆菌酶的稳定结合位点的行为与体内启动子的预期行为不一致。