Wendelburg B J, Marzluff W F
Department of Chemistry, Florida State University, Tallahassee 32306.
Mol Cell Biol. 1992 Sep;12(9):4132-41. doi: 10.1128/mcb.12.9.4132-4141.1992.
The formation of the 3' end of vertebrate small nuclear RNAs (snRNAs) requires that transcription initiate from an snRNA promoter. There is a loosely conserved required box 5 to 20 nucleotides (nt) 3' of the gene. The sea urchin snRNA genes contain promoter elements different from those of the vertebrate snRNAs. They also contain a characteristic 3' 15-nt sequence which is conserved among different sea urchin snRNA genes. We used microinjection of sea urchin U1 snRNA genes into sea urchin zygotes to define the sequence requirements for U1 snRNA 3'-end formation. Surprisingly, the conserved 3' box is not required for efficient 3'-end formation in vivo. Deletion analysis reveals that the 6 nt immediately 3' of the U1 snRNA are involved in 3'-end formation. Substitution analysis revealed that either these 6 nt 3' of the U1 RNA or the conserved 3' box could direct 3'-end formation. Transcripts initiated from a histone H4 promoter formed U1 3' ends about 50% as efficiently as transcripts initiated from the U1 promoter, even when the U1 end was placed in tandem with a histone 3'-processing signal, suggesting that transcription from an snRNA promoter is not necessary for formation of the 3' end of sea urchin U1 snRNA.
脊椎动物小核RNA(snRNA)3'末端的形成要求转录从snRNA启动子起始。在基因的3'端存在一个位置大致保守的、位于基因下游5至20个核苷酸(nt)处的必需框。海胆snRNA基因包含与脊椎动物snRNA不同的启动子元件。它们还含有一个特征性的3'端15个核苷酸序列,该序列在不同的海胆snRNA基因中是保守的。我们将海胆U1 snRNA基因显微注射到海胆受精卵中,以确定U1 snRNA 3'末端形成的序列要求。令人惊讶的是,保守的3'框在体内高效3'末端形成过程中并非必需。缺失分析表明,U1 snRNA紧邻3'端的6个核苷酸参与3'末端形成。替换分析显示,U1 RNA 3'端的这6个核苷酸或保守的3'框均可指导3'末端形成。从组蛋白H4启动子起始的转录本形成U1 3'末端的效率约为从U1启动子起始的转录本的50%,即使U1末端与组蛋白3'加工信号串联排列时也是如此,这表明对于海胆U1 snRNA 3'末端的形成而言,从snRNA启动子转录并非必需。