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首页> 外文期刊>Molecular and Cellular Biology >Effect of 5' splice site mutations on splicing of the preceding intron.
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Effect of 5' splice site mutations on splicing of the preceding intron.

机译：5'剪接位点突变对前面的内含子剪接的影响。

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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 and of the exon supports the suggestion that exons, rather than splice sites, are the recognition units for assembly of the spliceosome.
机译：含有相同内含子和外显子序列的三个外显子构建体在以内含子2开始的5'剪接位点发生突变，并在体外测定了突变对上游内含子剪接的影响。通过定量剪接产物或释放的套索状RNA确定，在5'剪接位点内两个或六个碱基的改变使内含子1的去除减少了至少20倍。突出的产物是将外显子1剪接至外显子3。检查复合物的形成表明，终止外显子2的5'剪接位点的突变降低了仅含有受影响外显子的前体RNA在体外直接装配的能力。这些结果表明内部外显子末端的突变抑制了外显子被剪接因子识别的能力。比较已知突变体位于内部外显子末端的脊椎动物5'剪接位点突变，表明外显子跳跃是该类型突变的优选表型，与此处报道的体外观察结果一致。通过在远端和外显子处的突变来抑制剪接支持了这样的建议，即外显子而不是剪接位点是用于组装剪接体的识别单元。

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