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细胞核中的RNA代谢:骨髓瘤细胞核中κ外显子的选择性转运以及感染的HeLa细胞核中腺病毒转录本的选择性转运。

RNA metabolism in nuclei: selective transport of kappa exons from myeloma nuclei and adenoviral transcripts from infected HeLa nuclei.

作者信息

Patterson R, Werner E, Fetherston J

出版信息

Nucleic Acids Res. 1986 May 27;14(10):4159-69. doi: 10.1093/nar/14.10.4159.

Abstract

Two independent systems and several analytical procedures have been used to establish that isolated mammalian nuclei selectively transport mature RNA polymerase I and II products. Murine myeloma nuclei retain physiologic restriction in our transport assay as assessed by the transport of the immunoglobulin kappa light chain mRNA and 18S and 28S rRNAs. Nearly 50% of the total kappa exons are transported as structurally intact mature mRNA molecules while less than 8% of either pulse-labeled or steady state kappa intron sequences are detected in the transported fraction. Ribosomal external transcribed spacer sequences also are absent in transported RNA. Release of cytoplasmic RNA from the outer nuclear membrane during the transport assay accounts for less than 10% of transported RNA. Nuclei isolated from adenovirus-infected HeLa cells at 20 hours post infection retain cellular actin mRNA and transport viral poly A+RNA. Ribosomal RNA is transported from infected nuclei although at a reduced rate compared to transport from mock-infected nuclei. Inhibition of transport of host mRNA is paralleled by the absence of pulse-labeled actin mRNA in the cytoplasm of infected cells. The implications of our transport data in relationship to intranuclear RNA trafficking are discussed.

摘要

已经使用了两个独立的系统和几种分析程序来确定分离的哺乳动物细胞核选择性地转运成熟的RNA聚合酶I和II产物。通过免疫球蛋白κ轻链mRNA以及18S和28S rRNA的转运评估,小鼠骨髓瘤细胞核在我们的转运试验中保持生理限制。将近50%的总κ外显子作为结构完整的成熟mRNA分子被转运,而在转运部分中检测到的脉冲标记或稳态κ内含子序列不到8%。转运的RNA中也不存在核糖体外部转录间隔序列。在转运试验期间,从外核膜释放的细胞质RNA占转运RNA的比例不到10%。感染后20小时从腺病毒感染的HeLa细胞中分离的细胞核保留细胞肌动蛋白mRNA并转运病毒多聚腺苷酸化RNA。核糖体RNA从感染的细胞核中转运出来,尽管与从 mock 感染的细胞核转运相比速率降低。宿主mRNA转运的抑制与感染细胞细胞质中不存在脉冲标记的肌动蛋白mRNA平行。讨论了我们的转运数据与核内RNA运输相关的意义。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4159/339852/01e8db4b7419/nar00279-0190-a.jpg

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