Sergeant A, Tigges M A, Raskas H J
J Virol. 1979 Mar;29(3):888-98. doi: 10.1128/JVI.29.3.888-898.1979.
The intranuclear structure of parental adenovirus 2 DNA was studied using digestion with micrococcal nuclease as a probe. When cultures were infected with 32P-labeled virions, at a multiplicity of 3,000 particles per cell, 14 to 21% of parental DNA penetrated the cell and reached the nucleus. Of this parental DNA, 60% could be solubilized by extensive digestion with micrococcal nuclease. The nuclease-resistant fraction contained viral deoxyribonucleoprotein monomers and oligomers. These nucleosome-like structures contained DNA fragments which are integral multiples of a unit-length DNA of approximately 185 base pairs. The monomeric DNA is similar in length to the unit-length DNA contained in cellular nucleosomes. However, the viral oligomers are slightly smaller than their cellular counterparts. DNA-DNA hybridization demonstrated that all segments of the viral genome, including those expressed as mRNA only at late times, are represented in the nucleosomal viral DNA. The amount of early intranuclear viral chromatin was proportional to multiplicity of infection up to multiplicities of 4,000 particles per cell. However, viral transcriptional activity did not increase in direct proportion to the amount of viral chromatin. Maximum accumulation of intranuclear viral chromatin was achieved by 3 h after infection. The intranuclear parental viral chromatin remained resistant to nuclease digestion even at late times in infection, after viral DNA replication had begun.
以微球菌核酸酶消化作为探针，研究了亲代腺病毒2型DNA的核内结构。当用32P标记的病毒粒子感染细胞培养物时，感染复数为每细胞3000个粒子，14%至21%的亲代DNA穿透细胞并到达细胞核。在这些亲代DNA中，60%可通过用微球菌核酸酶进行广泛消化而溶解。核酸酶抗性部分包含病毒脱氧核糖核蛋白单体和寡聚体。这些核小体样结构包含的DNA片段是约185个碱基对的单位长度DNA的整数倍。单体DNA的长度与细胞内核小体中所含的单位长度DNA相似。然而，病毒寡聚体比其细胞对应物略小。DNA-DNA杂交表明，病毒基因组的所有片段，包括那些仅在晚期表达为mRNA的片段，都存在于核小体病毒DNA中。早期核内病毒染色质的量与感染复数成正比，直至每细胞4000个粒子的感染复数。然而，病毒转录活性并未与病毒染色质的量成正比增加。感染后3小时达到核内病毒染色质的最大积累量。即使在感染后期病毒DNA复制开始后，核内亲代病毒染色质对核酸酶消化仍具有抗性。
