SEQUENCE REQUIREMENTS FOR SPLICING OF HIGHER EUKARYOTIC NUCLEAR PRE-MESSENGER-RNA
SEQUENCE REQUIREMENTS FOR SPLICING OF HIGHER EUKARYOTIC NUCLEAR PRE-MESSENGER-RNA
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DOI：
10.1016/0092-8674(86)90620-3
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发表时间：
1986-11-21
期刊：
影响因子：
64.5
通讯作者：
WEISSMANN, C
中科院分区：
文献类型：
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作者：
AEBI, M;HORNIG, H;WEISSMANN, C
我们确定了大兔β-的5”和3”剪接区中24个点突变对剪接的影响珠蛋白内含子在体外，3“AG突变显著减少5”切割并消除剪接。在体内，相同的突变在隐蔽的而不是正确的3“剪接位点引起有效的剪接。在体外，共有5“剪接区除了2个位置以外的所有位置的突变损害正确剪接并促进外显子1与外显子3的连接。在体内，除了将5“GT转化为AT或GA的那些突变外，相同的突变没有显示出任何效果，这些突变在体外和体内引起乳酸中间体的积累。我们的结论是，5“GT不需要为5”切割保守，它在3“剪接位点的切割和外显子连接中起重要作用。
We determined the effect on splicing of 24 point mutations in the 5'' and 3'' splice region of the large rabbit .beta.-globin intron. In vitro, 3'' AG mutations drastically reduced 5'' cleavage and abolish splicing. In vivo, the same mutations elicit efficient splicing at a cryptic, rather than the correct, 3'' splice site. In vitro, mutations at all but 2 positions of the consensus 5'' splice region impair correct splicing and promote joining of exon 1 to exon 3. In vivo, the same mutations show no effect, except for those converting 5'' GT to AT or GA, which cause accumulation of lariat intermediate in vitro and in vivo. We conclude that the 5'' GT need not be conserved for 5'' cleavage and that it plays an important role in cleavage and exon joining at the 3'' splice site.