Segregation of viral double‐stranded and single‐stranded DNA molecules in nucleo fo adenovirus infected cells as revealed by electron microscope in situ hybridization
Segregation of viral double‐stranded and single‐stranded DNA molecules in nucleo fo adenovirus infected cells as revealed by electron microscope in situ hybridization
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电子显微镜原位杂交显示核病毒感染细胞中病毒双链和单链 DNA 分子的分离
DOI：
10.1016/0248-4900(92)90206-g
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发表时间：
1992
影响因子：
2.7
通讯作者：
E. Pichard
中科院分区：
文献类型：
--
作者：
F. Puvion;E. Pichard
通过原位杂交技术在超微结构水平上研究了5型腺病毒感染的HeLa细胞核中子代病毒的形成，该技术允许特异性检测病毒双链DNA（dsDNA）或单链DNA（ssDNA）。在病毒基因组开始复制之前，进入靶细胞核的感染性DNA分子随机分布在宿主染色质纤维中，包括核仁相关染色质。它们是双链的，也就是说，没有单链断裂。病毒DNA与宿主浓缩染色质的这种结合也发生在有丝分裂中。病毒基因组复制的起始与核质中含有混合的病毒dsDNA和ssDNA的小纤维区域的出现同时发生。后来，在核转化的中间阶段，病毒dsDNA和ssDNA分子几乎完全分离成两个连续的亚结构。在此阶段，偶尔在病毒ssDNA积累位点附近观察到病毒。再后来，在细胞核中心形成了一个额外的亚结构，由大量的病毒dsDNA、微量的病毒ssDNA和大量的病毒组成。部分病毒ssDNA甚至在核转化的晚期阶段附着于一些病毒，这种关联强烈表明至少一些病毒基因组在它们仍在进行复制时发生了重排。
Formation of progeny viruses in the nuclei of HeLa cells infected with adenovirus type 5 was studied at the ultrastructural level byin situhybridization techniques allowing specific detection of either viral double‐stranded DNA (dsDNA) or single‐stranded DNA (ssDNA). Prior to the initiation of replication of viral genomes, infective DNA molecules which entered the nucleus of the target cell were randomly distributed among host chromatin fibers including nucleolus‐associated chromatin. They were double‐stranded, that is, without single‐strand breaks. Such association of viral DNA with host condensed chromatin also occurred in mitosis. The initiation of viral genome replication occufred simultaneously with the appearance in the nucleoplasm of small fibrillar regions containing intermingled viral dsDNA and ssDNA. Later, at the intermediate stage of nuclear transformation, viral dsDNA and ssDNA molecules were almost entirely separated into two contiguous substructures. At this stage, viruses were observed occasionally in the vicinity of viral ssDNA accumulation sites. Still later, an additional substructure developed in the centre of the nucleus which consisted of large quantities of viral dsDNA, traces of viral ssDNA and abundant viruses. Portions of viral ssDNA were attached to some viruses even at late stage of nuclear transformation, an association which strongly suggests the occurence of encapsidation of at least some of the viral genomes while they are still engaged in replication.