Effect of 5' splice site mutations on splicing of the preceding intron.
Effect of 5' splice site mutations on splicing of the preceding intron.
复制标题
5剪接位点突变对前面内含子剪接的影响。
DOI：
10.1128/mcb.10.12.6299-6305.1990
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发表时间：
1990
影响因子：
5.3
通讯作者：
Berget,SM
中科院分区：
文献类型：
--
作者：
Talerico,M;Berget,SM
将含有相同内含子和外显子序列的三个外显子构建体在内含子2开始的5 '剪接位点处进行突变，并在体外分析突变对上游内含子剪接的影响。5′剪接位点内2个或6个碱基的改变使内含子1的去除减少至少20倍，这通过对剪接产物或释放的IRNA进行定量来确定。突出的产物是外显子1到外显子3的跳跃剪接。复合物形成的检查表明，终止外显子2的5′剪接位点的突变抑制了仅含有受影响外显子的前体RNA在体外直接组装的能力。这些结果表明，突变的内部外显子的末端抑制的能力，外显子被识别的剪接因子。已知脊椎动物5′剪接位点突变（突变位于内部外显子末端）的比较表明，外显子跳跃是这种类型突变的首选表型，与本文报道的体外观察结果一致。外显子远端突变抑制剪接支持了外显子而不是剪接位点是剪接体组装的识别单位的观点。
Three exon constructs containing identical intron and exon sequences were mutated at the 5′ splice site beginning intron 2 and assayed for the effect of the mutation on splicing of the upstream intron in vitro. Alteration of two or six bases within the 5′ splice site reduced removal of intron 1 at least 20-fold, as determined by quantitation of either spliced product or released lariat RNA. The prominent product was skip splicing of exon 1 to exon 3. Examination of complex formation indicated that mutation of the 5′ splice site terminating exon 2 depressed the ability of precursor RNAs containing just the affected exon to direct assembly in vitro. These results suggest that mutation at the end of an internal exon inhibits the ability of the exon to be recognized by splicing factors. A comparison of the known vertebrate 5′ splice site mutations in which the mutation resides at the end of an internal exon indicated that exon skipping is the preferred phenotype for this type of mutation, in agreement with the in vitro observation reported here. Inhibition of splicing by mutation at the distal end of the exon supports the suggestion that exons, rather than splice sites, are the recognition units for assembly of the spliceosome.