Zhuang Y, Weiner A M
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, Connecticut 06510.
Genes Dev. 1989 Oct;3(10):1545-52. doi: 10.1101/gad.3.10.1545.
We have developed an assay to test whether U2 snRNA can base-pair with the branch site during mammalian mRNA splicing. The beta 110 point mutation (GG----AG) within the first intron of human beta-globin generates a new 3' splice site that is preferentially used. We show here that use of the normal 3' splice site can be restored either by improving the match of a cryptic branch site to the branch site consensus or by introducing mutant U2 snRNAs with greater complementarity to the cryptic branch site. These data indicate that human U2 snRNA can form base pairs with the mRNA precursor; however, base pairing appears to be optional because some mammalian branch sites do not match the consensus.
我们开发了一种检测方法,以测试在哺乳动物mRNA剪接过程中U2 snRNA是否能与分支位点形成碱基对。人类β-珠蛋白第一个内含子内的β110点突变(GG----AG)产生了一个优先使用的新3'剪接位点。我们在此表明,通过改善隐蔽分支位点与分支位点共有序列的匹配度,或者通过引入与隐蔽分支位点具有更高互补性的突变U2 snRNAs,正常3'剪接位点的使用可以得到恢复。这些数据表明,人类U2 snRNA可以与mRNA前体形成碱基对;然而,碱基配对似乎是可有可无的,因为一些哺乳动物分支位点并不符合共有序列。