Bratosin S, Laub O, Tal J, Aloni Y
Proc Natl Acad Sci U S A. 1979 Sep;76(9):4289-93. doi: 10.1073/pnas.76.9.4289.
During an electron-microscopic survey with the aim of identifying the parvovirus MVM transcription template, we observed previously unidentified structures of MVM DNA in lysates of virus-infected cells. These included double-stranded "lasso"-like structures and relaxed circles. Both structures were of unit length MVM DNA, indicating that they were not intermediates formed during replication; they each represented about 5% of the total nuclear MVM DNA. The proportion of these structures was unchanged after digestion with sodium dodecyl sulfate/Pronase and RNase and after mild denaturation treatment. Cleavage of the "lasso" structures with EcoRI restriction endonuclease indicated that the "noose" part of the "lasso" structure is located on the 5' side of the genomic single-stranded MVM DNA. A model is presented for the molecular nature of the circularization process of MVM DNA in which the "lasso" structures are identified as intermediates during circle formation. This model proposes a mechanism for circularization of linear DNAs.
在一项旨在鉴定细小病毒MVM转录模板的电子显微镜调查中,我们在病毒感染细胞的裂解物中观察到了以前未鉴定出的MVM DNA结构。这些结构包括双链“套索”样结构和松弛环。两种结构均为单位长度的MVM DNA,这表明它们不是复制过程中形成的中间体;它们各自约占细胞核MVM DNA总量的5%。在用十二烷基硫酸钠/链霉蛋白酶和核糖核酸酶消化以及轻度变性处理后,这些结构的比例没有变化。用EcoRI限制性内切酶切割“套索”结构表明,“套索”结构的“套索”部分位于基因组单链MVM DNA的5'侧。本文提出了一个关于MVM DNA环化过程分子本质的模型,其中“套索”结构被确定为环化过程中的中间体。该模型提出了线性DNA环化的机制。