INTERVENING SEQUENCES INCREASE EFFICIENCY OF RNA 3' PROCESSING AND ACCUMULATION OF CYTOPLASMIC RNA
INTERVENING SEQUENCES INCREASE EFFICIENCY OF RNA 3' PROCESSING AND ACCUMULATION OF CYTOPLASMIC RNA
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DOI：
10.1093/nar/18.4.937
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发表时间：
1990-02-25
影响因子：
14.9
通讯作者：
GORMAN, CM
中科院分区：
文献类型：
--
作者：
HUANG, MTF;GORMAN, CM
构建了两种表达载体，其不同之处仅在于它们的5“-非翻译前导序列中存在（+）或不存在（-）间插序列（IVS）。瞬时转染到四种哺乳动物细胞系中导致来自IVS（+）载体的指示蛋白（CAT）水平更高（6至50=倍）。在293 s和HeLa细胞系的细胞质RNA浓度直接对应于所得的蛋白质水平;在293 s细胞的转录起始和延伸，稳态总核RNA，和细胞质RNA的稳定性的测量，是等效的两个载体。令人惊讶的是，来自IVS（+）载体的poly（A）+核RNA的量更大。由于这种差异与多聚腺苷酸化的细胞质RNA的比例相匹配，我们的结果意味着剪接与多聚腺苷酸化/转运途径相结合。
Two expression vectors were constructed that differ only in the presence (+) or absence (-) of an intervening sequence (IVS) in their 5''-untranslated leaders. Transient transfection into four mammalian cell lines resulted in higher levels of the indicator protein (CAT) from the IVS(+) vector (6 to 50=fold). Cytoplasmic RNA concentrations in 293s and HeLa cell lines corresponded directly to resultant protein levels; measurements in 293s cells of transcription initiation and elongation, steady-state total nuclear RNA, and cytoplasmic RNA stability, were equivalent for the two vectors. Surprisingly, the amount of poly(A)+ nuclear RNA was greater from the IVS(+) vector. Since this difference matches the ratio seen with polyadenylated cytoplasmic RNA, our results imply that splicing is coupled to a polyadenylation/transport pathway.