Yaneva J, Zlatanova J, Paneva E, Srebreva L, Tsanev R
Institute of Molecular Biology, Bulgarian Academy of Science, Sofia.
FEBS Lett. 1990 Apr 24;263(2):225-8. doi: 10.1016/0014-5793(90)81379-3.
By using direct competition experiments, the binding of histone H1AB (a mixture of H1A and H1B) and H1(0) to superhelical and linear DNA forms was studied. Mouse liver H1 isohistones and plasmid p alpha GD containing part of the 5' flanking and part of the coding sequence of the mouse alpha-globin gene in pUC18 were used as partners in the binding reaction. The competition experiments were performed by direct mixing of the histone with labelled supercoiled DNA (at 125 mM NaCl and at a histone/DNA ratio of 1.0) and addition to the mixture of increasing amounts of cold competitor DNA, either supercoiled or linear. The radioactivity of the complex formed was determined by filter binding. The results show that both histones H1 and H1(0) posses a strong binding preference for supercoiled DNA forms. Thus, histone H1(0) resembles the regular somatic set of histone H1 and not the other differentiation-specific histone H5 studied thus far.
通过直接竞争实验,研究了组蛋白H1AB(H1A和H1B的混合物)和H1(0)与超螺旋和线性DNA形式的结合。小鼠肝脏H1同工组蛋白和包含小鼠α-珠蛋白基因5'侧翼部分和编码序列部分的质粒pαGD(位于pUC18中)被用作结合反应的搭档。竞争实验通过将组蛋白与标记的超螺旋DNA直接混合(在125 mM NaCl下,组蛋白/DNA比例为1.0),并向混合物中添加越来越多的冷竞争DNA(超螺旋或线性)来进行。形成的复合物的放射性通过滤膜结合来测定。结果表明,组蛋白H1和H1(0)都对超螺旋DNA形式具有强烈的结合偏好。因此,组蛋白H1(0)类似于常规的体细胞组蛋白H1,而不像迄今为止研究的其他分化特异性组蛋白H5。