Soltyk A, Shugar D, Piechowska M
J Bacteriol. 1975 Dec;124(3):1429-38. doi: 10.1128/jb.124.3.1429-1438.1975.
With competent cultures of Bacillus subtilis the uptake of Escherichia coli deoxyribonucleic acid (DNA) is about 50% that for homologous DNA. Uptake of phage T6 DNA, if any, is of the order of 7%, while nonglucosylated phage T6 (T6) DNA is taken up almost as effectively as homologous DNA. Both T6 and T4 DNA interfere only minimally with uptake of homologous DNA; by contrast, T6 DNA competes with homologous DNA as effectively as the latter itself. These results indicate that the glucose residues in the T-even phage DNA, located in the large groove of the DNA helix, reduce affinity for cellular receptors, leading to low binding of T6 DNA. The latter DNA is considerably less degraded by extracellular nucleases than homologous DNA, thus excluding enzymatic hydrolysis as the source of poor uptake. Affinity of DNA for competent cells was also evaluated by the formation, and detection in a CsCl density gradient, of complexes of DNA with cellular constituent(s). Such comlexes, similar to those previously observed with transforming DNA, are formed by E. coli DNA and T6 DNA; in reconstruction experiments the denatured forms of these same DNA samples form complexes when added to the cells before lysis. T6 DNA, on the other hand, does not form such a complex. The possible role of such complexes in transport of DNA to the cell interior is discussed.
对于枯草芽孢杆菌感受态培养物而言,大肠杆菌脱氧核糖核酸(DNA)的摄取量约为同源DNA摄取量的50%。噬菌体T6 DNA的摄取量(如果有的话)约为7%,而未糖基化的噬菌体T6(T6)DNA的摄取效率几乎与同源DNA相同。T6和T4 DNA对同源DNA摄取的干扰都很小;相比之下,T6 DNA与同源DNA的竞争效果与同源DNA自身相当。这些结果表明,位于DNA螺旋大沟中的T偶数噬菌体DNA中的葡萄糖残基降低了对细胞受体的亲和力,导致T6 DNA的低结合。后一种DNA被细胞外核酸酶降解的程度远低于同源DNA,因此排除了酶促水解是摄取不佳的原因。还通过在CsCl密度梯度中形成并检测DNA与细胞成分的复合物来评估DNA对感受态细胞的亲和力。这种复合物类似于先前观察到的转化DNA形成的复合物,由大肠杆菌DNA和T6 DNA形成;在重建实验中,这些相同DNA样品的变性形式在细胞裂解前加入时会形成复合物。另一方面,T6 DNA不会形成这样的复合物。文中讨论了这种复合物在DNA转运到细胞内部过程中可能发挥的作用。