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首页> 外文期刊>Molecular and Cellular Biology >Factors influencing alternative splice site utilization in vivo.
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Factors influencing alternative splice site utilization in vivo.

机译：影响体内替代剪接位点利用的因素。

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To study factors that influence the choice of alternative pre-mRNA splicing pathways, we introduced plasmids expressing either wild-type or mutated simian virus 40 (SV40) early regions into tissue culture cells and then measured the quantities of small-t and large-T RNAs produced. One important element controlling splice site selection was found to be the size of the intron removed in the production of small-t mRNA; expansion of this intron (from 66 to 77 or more nucleotides) resulted in a substantial increase in the amount of small-t mRNA produced relative to large-T mRNA. This suggests that in the normal course of SV40 early pre-mRNA processing, large-T splicing is at a competitive advantage relative to small-t splicing because of the small size of the latter intron. Several additional features of the pre-mRNA that can influence splice site selection were also identified by analyzing the effects of mutations containing splice site duplications. These include the strengths of competing 5' splice sites and the relative positions of splice sites in the pre-mRNA. Finally, we showed that the ratio of small-t to large-T mRNA was 10 to 15-fold greater in human 293 cells than in HeLa cells or other mammalian cell types. These results suggest the existence of cell-specific trans-acting factors that can dramatically alter the pattern of splice site selection in a pre-mRNA.
机译：为了研究影响替代前mRNA剪接途径选择的因素，我们将表达野生型或突变猿猴病毒40（SV40）早期区域的质粒引入组织培养细胞中，然后测量了小T和大T的数量产生的RNA。发现控制剪接位点选择的一个重要因素是在生产小t mRNA时去除的内含子大小。内含子的扩增（从66个核苷酸增加到77个或更多个核苷酸）导致产生的小-t mRNA的量相对于大-t mRNA显着增加。这表明在SV40早期pre-mRNA加工的正常过程中，大T剪接相对于小t剪接具有竞争优势，因为后者内含子的大小较小。通过分析含有剪接位点重复的突变的影响，还确定了前mRNA可以影响剪接位点选择的其他几个特征。这些包括竞争性5'剪接位点的强度以及pre-mRNA中剪接位点的相对位置。最后，我们显示，人293细胞中的小T与大T mRNA的比率比HeLa细胞或其他哺乳动物细胞类型大10至15倍。这些结果表明存在细胞特异性反式作用因子，其可显着改变前mRNA中剪接位点选择的模式。

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