Besse S, Puvion-Dutilleul F
Institut Fédératif CNRS, UPR 9044, Laboratoire Organisation fonctionnelle du noyau, Villejuif, France.
Eur J Cell Biol. 1996 Sep;71(1):33-44.
Herpes simplex virus type 1 (HSV-1) infection of HeLa cells induces profound changes in the structure of the nucleoli. They become markedly elongated, and their fibrillar centers become greatly diminished in number, but larger than in non-infected HeLa cells, and only partially surrounded by the dense fibrillar component. The effect of prolonged HSV-1 infection on the distribution of the rRNA genes was studied by means of postembedding electron microscope in situ hybridization using a biotinylated ribosomal DNA (rDNA) probe, which spans about half of the rRNA gene, and subsequent immunogold labeling of the resulting hybrids. Gold particles accumulated over two structures: a large solitary, finely fibrillar, moderately electron opaque area which was detectable only in a few sections of nucleoli and corresponded to the virus-modified fibrillar center, and over limited areas of the nucleolus-associated chromatin. In non-infected HeLa cells, foci of clustered rRNA genes were observed in the more frequently detected fibrillar centers and in association with condensed chromatin. It would be expected that foci of extended rDNA molecules might contain active or potentially active genes, whereas foci of highly compacted rDNA molecules might contain inactive genes. The ribosomal RNA molecules which were detected with the same probe over the dense fibrillar component and the granular component of the nucleoli of both infected and non-infected cells were not found within the rDNA-containing foci. The data strongly suggest that the changes in the size and number of fibrillar centers induced by the intranuclear development of HSV-1 might be directly linked to the well-known decrease of the nucleolar activity.
单纯疱疹病毒1型(HSV-1)感染HeLa细胞会引起核仁结构的深刻变化。核仁明显拉长,其纤维中心数量大幅减少,但比未感染的HeLa细胞中的纤维中心更大,且仅部分被致密纤维成分包围。通过使用生物素化核糖体DNA(rDNA)探针进行包埋后电子显微镜原位杂交,研究了HSV-1长期感染对rRNA基因分布的影响,该探针跨越约一半的rRNA基因,随后对所得杂交体进行免疫金标记。金颗粒聚集在两种结构上:一个大的孤立的、细纤维状的、中等电子不透明区域,仅在少数核仁切片中可检测到,对应于病毒修饰的纤维中心,以及在核仁相关染色质的有限区域。在未感染的HeLa细胞中,在更频繁检测到的纤维中心以及与浓缩染色质相关的区域观察到成簇的rRNA基因位点。可以预期,延伸的rDNA分子位点可能包含活跃或潜在活跃的基因,而高度压缩的rDNA分子位点可能包含不活跃的基因。在感染和未感染细胞的核仁致密纤维成分和颗粒成分上用相同探针检测到的核糖体RNA分子,在含rDNA的位点内未发现。数据强烈表明,HSV-1在核内发育引起的纤维中心大小和数量的变化可能与核仁活性的众所周知的降低直接相关。