Hepatocarcinogen-induced alterations in nuclear RNA compartmentation.
Hepatocarcinogen-induced alterations in nuclear RNA compartmentation.
复制标题
肝癌诱导的核 RNA 区室变化。
DOI：
10.1093/carcin/8.9.1235
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发表时间：
1987
期刊：
影响因子：
4.7
通讯作者：
Woo,CH
中科院分区：
文献类型：
--
作者：
Clawson,GA;Button,JD;Woo,CH
纯化预标记的大鼠肝细胞核，用去污剂漂洗以去除细胞质 RNA 污染物，并在两个体外 RNA 转运测定中孵育。在含有聚乙烯吡咯烷酮 (PVP) 的测定中保持核 RNA 序列的适当划分，防止水介质中的核膨胀。在这些条件下，在存在PVP（和ATP）的情况下释放的poly(A)+RNA在指导体外蛋白质合成方面比在不存在任一添加剂的情况下释放的poly(A)+RNA显着更活跃。此外，α1-酸性糖蛋白的第五个内含子被切除后仍然受到细胞核限制，而在没有 PVP 的情况下会释放出聚（A）+加工分解代谢物。用肝癌致癌物硫代乙酰胺治疗大鼠后，核序列的适当区划丢失，与体内情况相似。该测定对于核细胞质 RNA 体外转运的后续研究很有用，特别是与癌发生起始相关的 RNA 转运改变相关的研究。
Prelabeled rat liver nuclei were purified, detergent-rinsed to remove cytoplasmic RNA contaminants and incubated in twoin vitroRNA transport assays. Appropriate compartmentation of nuclear RNA sequences was maintained in an assay containing polyvinylpyrrolidone (PVP), which prevents nuclear swelling in aqueous media. Under these conditions, poly (A)+RNA released in the presence of PVP (and ATP) was significantly more active in directingin vitroprotein synthesis than poly(A)+RNA released in the absence of either additive. Further, the excised fifth intron ofα1-acid glycoprotein remained nucleus-restricted, while release of poly(A)+processing catabolites occurred in the absence of PVP. The appropriate compartmentation of nuclear sequences was lost following treament of rats with the hepato-carcinogen thioacetamide, paralleling thein vivosituation. This assay should prove useful for subsequent studies on nucleocytoplasmic RNA transportin vitro, in particular for investigations relating to the altered RNA transport associated with the initiation of carcinogenesis.