Hwan J C, Leffak I M, Li H J, Huang P C, Mura C
Biochemistry. 1975 Apr 8;14(7):1390-6. doi: 10.1021/bi00678a008.
Histone V (2fc) from chick erythroctes was used in the study of its interaction with DNA from various sources. Complexes between this histone and DNA were formed using the procedure of continuous NaCl gradient dialysis in urea. Two physical methods, namely thermal denaturation and circular dichroism (CD), were used as analytical tools. Thermal denaturation of nucleohistone V with chick or calf thymus DNA shows three melting bands: band I at 45-50 degrees corresponds to free base pairs; band II at 75-79 degrees, and band III at 90-93 degrees correspond to histone-bound base pairs. In histone-bound regions, there are 1.5 amino acid residues/nucleotide in nucleohistone V. In contrast, a value between 2.9 and 3.3 was determined for nucleohistone I (fl) (H. J. Li (1973), Biopolymers 12, 287). Similar melting properties have been observed for histone V complexed with bacterial DNA from Micrococcus luteus. Histone V binding to DNA induces a slight transition from a B-type CD spectrum to a C-type spectrum. Trypsin treatment of nucleohistone V reduces melting band III much more effectively than band II. Such a treatment also restores DNA to B conformation in the free state. Reduction of the melting bands of nucleohistone V by polylysine binding follows the order of I greater than II greater than III, accompanied by the increase of a new band at 100 degrees. When two bacterial DNAs of varied A + T (adenine + thymine) content simultaneously compete for the binding of histone V, the more (A " T)-rich DNA is selectively favored. Under experimental conditions described here, Clostridium perfringens DNA with 69% A + T is bound by histone V in preference to chicken DNA with 56% A + T although the latter has natural sequences for histone V binding.
来自鸡红细胞的组蛋白V(2fc)被用于研究其与各种来源DNA的相互作用。使用尿素中连续NaCl梯度透析的方法形成了这种组蛋白与DNA之间的复合物。两种物理方法,即热变性和圆二色性(CD),被用作分析工具。组蛋白V与鸡或小牛胸腺DNA形成的核组蛋白的热变性显示出三个解链带:45 - 50摄氏度的带I对应于游离碱基对;75 - 79摄氏度的带II和90 - 93摄氏度的带III对应于与组蛋白结合的碱基对。在与组蛋白结合的区域,核组蛋白V中每核苷酸有1.5个氨基酸残基。相比之下,核组蛋白I(fl)的该值在2.9至3.3之间(H. J. 李(1973年),《生物聚合物》12,287)。对于与藤黄微球菌的细菌DNA复合的组蛋白V,也观察到了类似的解链特性。组蛋白V与DNA的结合会导致从B型CD光谱轻微转变为C型光谱。用胰蛋白酶处理核组蛋白V时,带III的解链比带II更有效。这种处理还能使游离状态的DNA恢复到B构象。聚赖氨酸结合使核组蛋白V的解链带减少的顺序为I > II > III,同时在100摄氏度出现一个新带。当两种A + T(腺嘌呤 + 胸腺嘧啶)含量不同的细菌DNA同时竞争组蛋白V的结合时,富含(A + T)的DNA更受青睐。在此处描述的实验条件下,尽管鸡DNA具有组蛋白V结合的天然序列,但含有69% A + T的产气荚膜梭菌DNA比含有56% A + T的鸡DNA更优先被组蛋白V结合。