5-Fluorouracil substitution alters pre-mRNA splicing in vitro.
5-Fluorouracil substitution alters pre-mRNA splicing in vitro.
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DOI：
10.1016/s0021-9258(18)68949-5
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发表时间：
1988-03
期刊：
影响因子：
--
通讯作者：
S. Doong;B. Dolnick
中科院分区：
文献类型：
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作者：
S. Doong;B. Dolnick
我们感兴趣的是确定药物5-氟尿嘧啶掺入前mRNA是否可以改变RNA加工。研究了5-氟尿嘧啶（FUra）取代对前体mRNA体外剪接的影响。在UTP、FUTP或两者存在下合成含有前两个外显子和第一个间插序列的32 P标记的人β-珠蛋白前mRNA。在体外剪接反应产生了几个异常的中间体和产品从FURa取代的成绩单。新的次要剪接产物的出现取决于剪接反应的pH和FUra掺入前体mRNA的程度。在pH 8.4和7.7下观察到异常剪接，但在pH 6.7下未观察到异常剪接。对新的次要剪接产物进行测序，发现其含有来自间插序列3 '端的额外20个碱基。异常迁移中间体具有与正常lariat-exon中间体相同的结构。这些结果表明，将FUra取代到前体mRNA中可以改变体外剪接。
We are interested in determining whether incorporation of the drug 5-fluorouracil into pre-mRNA can alter RNA processing. The effect of 5-fluorouracil (FUra) substitution on the in vitro splicing of pre-mRNA was studied. 32P-Labeled human beta-globin pre-mRNA containing the first two exons and the first intervening sequence was synthesized in the presence of UTP, FUTP, or both. In vitro splicing reactions generated several abnormal intermediates and products from FUra-substituted transcripts. The appearance of a new minor spliced product was dependent on both the pH of the splicing reaction and the extent of FUra incorporation into pre-mRNA. Abnormal splicing was observed at pH 8.4 and 7.7 but not at pH 6.7. The new minor spliced product was sequenced and found to contain an additional 20 bases derived from the 3'-end of the intervening sequence. The abnormally migrating intermediate had the same structure as the normal lariat-exon intermediate. These results suggest that FUra substitution into pre-mRNA can alter splicing in vitro.