Dymshits G M, Fet V Ia
Mol Biol (Mosk). 1977 May-Jun;11(3):531-6.
In DNA preparations isolated from regenerating rat liver 22 hours after partial hepatectomy, i.e. at the period of the most intensive DNA synthesis a "Denaturating Protein Factor" (DPF) tightly bound to DNA was found. Isolated protein fraction with a molecular weight of 6500 dalton was found to be homogenous upon SDS-polyacrylamide electrophoresis. The degree of destabilisation of DNA was estimated by its reaction with water-soluble [14C]CME-carbodiimide which modifies selectively guanine and thymine residues only in the denatured DNA regions. Pronase treated DPF loses its DNA-denaturing capacity. Pronase treatment of DNA--DPF complex restores native DNA structure. DPF from rat liver was able to denature DNA from calf thymus and bacteriophage T7 DNA. A hypothesis is proposed that the DPF is responsible for the destabilization of DNA secondary structure in the process of replication.
在部分肝切除术后22小时从再生大鼠肝脏中分离得到的DNA制剂中,即在DNA合成最活跃的时期,发现了一种与DNA紧密结合的“变性蛋白因子”(DPF)。经SDS - 聚丙烯酰胺电泳分析,分离得到的分子量为6500道尔顿的蛋白质组分是均一的。通过DNA与水溶性[14C]CME - 碳二亚胺的反应来估计DNA的去稳定程度,该试剂仅选择性修饰变性DNA区域中的鸟嘌呤和胸腺嘧啶残基。经链霉蛋白酶处理的DPF失去其DNA变性能力。对DNA - DPF复合物进行链霉蛋白酶处理可恢复天然DNA结构。来自大鼠肝脏的DPF能够使小牛胸腺DNA和噬菌体T7 DNA变性。提出了一种假说,即DPF在复制过程中负责DNA二级结构的去稳定。