van Wielink P S, Naaktgeboren N, Sussenbach J S
Biochim Biophys Acta. 1979 Jun 20;563(1):89-99. doi: 10.1016/0005-2787(79)90010-8.
Adenovirus type 5 contains linear double-stranded DNA with protein covalently attached to the ends of the molecules. The presence of protein at the termini of intracellular viral DNA in adenovirus type 5-infected cells was investigated at different stages during the replication process. The intracellular viral DNA was isolated from the nuclei by lysis in 4 M guanidine hydrochloride. Electrophoresis on agarose gels of HsuI restriction enzyme fragments and sucrose gradient centrifugation were used to detect protein on intracellular viral DNA. After uncoating parental DNA still contains protein attached to the termini of the viral genome. Replicating and mature progeny viral DNA can also be isolated in the form of DNA-protein complexes. These complexes exhibit the same properties as the DNA-protein complex isolated from purified virions. These results suggest that the protein at the termini of intracellular viral DNA is identical to the protein attached to the 5'-ends of the DNA extracted from virions and that it is possibly involved in the replication of viral DNA.
5型腺病毒含有线性双链DNA,其分子末端共价连接有蛋白质。在复制过程的不同阶段,对5型腺病毒感染细胞内病毒DNA末端蛋白质的存在情况进行了研究。通过在4M盐酸胍中裂解从细胞核中分离出细胞内病毒DNA。使用琼脂糖凝胶上的HsuI限制性酶切片段电泳和蔗糖梯度离心来检测细胞内病毒DNA上的蛋白质。脱壳后,亲本DNA仍含有附着在病毒基因组末端的蛋白质。复制的和成熟的子代病毒DNA也可以以DNA-蛋白质复合物的形式分离出来。这些复合物表现出与从纯化病毒粒子中分离出的DNA-蛋白质复合物相同的特性。这些结果表明,细胞内病毒DNA末端的蛋白质与从病毒粒子中提取的DNA 5'-末端附着的蛋白质相同,并且它可能参与病毒DNA的复制。