Bridge E, Riedel K U, Johansson B M, Pettersson U
Department of Medical Genetics, Uppsala University, Biomedical Center, Sweden.
J Cell Biol. 1996 Oct;135(2):303-14. doi: 10.1083/jcb.135.2.303.
Posttranscriptional steps in the production of mRNA include well characterized polyadenylation and splicing reactions, but it is also necessary to understand how RNA is transported within the nucleus from the site of its transcription to the nuclear pore, where it is translocated to the cytoplasmic compartment. Determining the localization of RNA within the nucleus is an important aspect of understanding RNA production and may provide clues for investigating the trafficking of RNA within the nucleus and the mechanism for its export to the cytoplasm. We have previously shown that late phase adenovirus-infected cells contain large clusters of snRNP and non-snRNP splicing factors; the presence of these structures is correlated with high levels of viral late gene transcription. The snRNP clusters correspond to enlarged interchromatin granules present in late phase infected cells. Here we show that polyadenylated RNA and spliced tripartite leader exons from the viral major late transcription unit are present in these same late phase snRNP-containing structures. We find that the majority of the steady state viral RNA present in the nucleus is spliced at the tripartite leader exons. Tripartite leader exons are efficiently exported from the nucleus at a time when we detect their accumulation in interchromatin granule clusters. Since the enlarged interchromatin granules contain spliced and polyadenylated RNA, we suggest that viral RNA may accumulate in this late phase structure during an intranuclear step in RNA transport.
信使核糖核酸(mRNA)产生过程中的转录后步骤包括特征明确的聚腺苷酸化和剪接反应,但了解RNA如何在细胞核内从转录位点转运至核孔(在核孔处它被转运至细胞质区室)也很有必要。确定RNA在细胞核内的定位是理解RNA产生的一个重要方面,并且可能为研究RNA在细胞核内的运输及其输出至细胞质的机制提供线索。我们之前已经表明,晚期腺病毒感染的细胞含有大量小核核糖核蛋白(snRNP)和非snRNP剪接因子簇;这些结构的存在与高水平的病毒晚期基因转录相关。snRNP簇对应于晚期感染细胞中出现的扩大的染色质间颗粒。在这里我们表明,来自病毒主要晚期转录单位的聚腺苷酸化RNA和剪接的三联前导外显子存在于这些相同的含有晚期snRNP的结构中。我们发现,细胞核中存在的大多数稳态病毒RNA在三联前导外显子处被剪接。当我们检测到三联前导外显子在染色质间颗粒簇中积累时,它们能有效地从细胞核输出。由于扩大的染色质间颗粒含有剪接的和聚腺苷酸化的RNA,我们认为病毒RNA可能在RNA运输的核内步骤中在这个晚期结构中积累。