The structure of replicating adenovirus DNA molecules: characterization of DNA-protein complexes from infected cells.
The structure of replicating adenovirus DNA molecules: characterization of DNA-protein complexes from infected cells.
复制标题
复制腺病毒 DNA 分子的结构：受感染细胞 DNA-蛋白质复合物的表征。
DOI：
10.1101/sqb.1979.043.01.080
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发表时间：
1979
期刊：
影响因子：
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通讯作者：
R. L. Lechner
中科院分区：
文献类型：
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作者：
T. J. Kelly;R. L. Lechner
腺病毒的基因组是分子量为20-29 × 106的非排列、线性、双链DNA分子（综述见Levine et al.1976）。从纯化的病毒颗粒中分离的成熟腺病毒（Ad）基因组已被证明含有两个新的结构特征。首先，Ad双链体的每条链的一端的核苷酸序列与另一端的序列互补。这种反向末端重复的存在首先从单位长度的Ad单链在杂交条件下环化的观察中推断出来（Wolfson和Dressler 1972; Garon等1972），并且最近通过直接序列分析证实了人2型腺病毒（Ad 2）和5型腺病毒（Ad 5（Steenbergh等人，1977年; JR Arrand和RJ Roberts，个人通讯）。第二，Ad链的5 '末端与蛋白质紧密结合（罗宾逊等1973;罗宾逊和Bellet 1975; Sharp等1976; Carusi 1977; Padmanabhan和Padmanabhan 1977; Rekosh等1977）。在Ad 2和Ad 5的情况下，末端蛋白具有55，000（55 k）的表观分子量，并且似乎通过共价键与DNA连接（Rekosh等人，1977）。在水溶液中，Ad基因组的末端通过其末端蛋白部分结合形成环状和寡聚体，这些环状和寡聚体可在电子显微镜中观察到（罗宾逊et al. 1973;罗宾逊and Bellett 1975; Sharp et al. 1976; Rekosh et al. 1977）。我们最近通过电子显微镜（EM）对从感染后20小时的KB细胞中分离的复制型Ad 2 DNA分子的结构进行了研究（Lechner和Kelly 1977）。分离程序包括一个步骤，其中复制分子通过用SDS和链霉蛋白酶处理脱蛋白，然后用苯酚提取。观察到两种基本类型的复制分子：具有一个或多个单链分支的Ad 2长度线性双链DNA分子（I型）和具有从一端延伸可变距离的单链区域的Ad 2长度线性DNA分子（II型）。一小部分分子具有
The genomes of the adenoviruses are nonpermuted, linear, duplex DNA molecules with molecular weights of 20-29 x 106 (for review, see Levine et al. 1976). Mature adenovirus (Ad) genomes isolated from purified virus particles have been shown to contain two novel structural features. First, the nucleotide sequence at one terminus of each strand of the Ad duplex is complementary to the sequence at the other terminus. The existence of this inverted terminal repetition was first inferred from the observation that unit-length Ad single strands cyclize when placed under hybridization conditions (Wolfson and Dressler 1972; Garon et al. 1972) and has been confirmed more recently for human adenovirus types 2 (Ad2) and 5 (Ad5) by direct sequence analysis (Steenbergh et al. 1977; JR Arrand and RJ Roberts, pers. comm.). Second, the 5'termini of Ad strands are tightly bound to a protein (Robinson et al. 1973; Robinson and Bellet 1975; Sharp et al. 1976; Carusi 1977; Padmanabhan and Padmanabhan 1977; Rekosh et al. 1977). In the cases of Ad2 and Ad5, the terminal protein has an apparent molecular weight of 55,000 (55k) and appears to be linked to the DNA by covalent bonds (Rekosh et al. 1977). In aqueous solution, the termini of Ad genomes associate via their terminal protein moities to form circles and oligomers which can be visualized in the electron microscope (Robinson et al. 1973; Robinson and Bellett 1975; Sharp et al. 1976; Rekosh et al. 1977). The functional roles of the terminal protein and the inverted terminal repetition in the Ad life cycle are not understood at present.We have recently carried out by electron microscopy (EM) a study of the structure of replicating Ad2 DNA molecules isolated from infected KB cells 20 hours after infection (Lechner and Kelly 1977). The isolation procedure included a step in which the replicating molecules were deproteinized by treatment with SDS and Pronase, followed by phenol extraction. Two basic types of replicating molecules were observed: Ad2-1ength linear duplex DNA molecules with one or more single-strand branches (type I) and Ad2-length linear DNA molecules with a single-strand region extending a variable distance from one end (type II). A small fraction of the molecules had the