Neuman de Vegvar H E, Dahlberg J E
Department of Physiological Chemistry, University of Wisconsin, Madison 53706.
Mol Cell Biol. 1990 Jul;10(7):3365-75. doi: 10.1128/mcb.10.7.3365-3375.1990.
We have analyzed the structures and locations of small nuclear RNA (snRNA) precursors at various stages in their synthesis and maturation. In the nuclei of pulse-labeled Xenopus laevis oocytes, we detected snRNAs that were longer than their mature forms at their 3' ends by up to 10 nucleotides. Analysis of the 5' caps of these RNAs and pulse-chase experiments showed that these nuclear snRNAs were precursors of the cytoplasmic pre-snRNAs that have been observed in the past. Synthesis of pre-snRNAs was not abolished by wheat germ agglutinin, which inhibits export of the pre-snRNAs from the nucleus, indicating that synthesis of these RNAs is not obligatorily coupled to their export. Newly synthesized U1 RNAs could be exported from the nucleus regardless of the length of the 3' extension, but pre-U1 RNAs that were elongated at their 3' ends by more than about 10 nucleotides were poor substrates for trimming in the cytoplasm. The structure at the 3' end was critical for subsequent transport of the RNA back to the nucleus. This requirement ensures that truncated and incompletely processed U1 RNAs are excluded from the nucleus.
我们分析了小核RNA(snRNA)前体在合成和成熟各个阶段的结构与位置。在经脉冲标记的非洲爪蟾卵母细胞核中,我们检测到3'端比其成熟形式长多达10个核苷酸的snRNA。对这些RNA的5'帽结构分析以及脉冲追踪实验表明,这些核snRNA是过去已观察到的细胞质前体snRNA的前体。麦胚凝集素抑制前体snRNA从细胞核输出,但前体snRNA的合成并未被其消除,这表明这些RNA的合成并非必然与其输出相偶联。新合成的U1 RNA无论3'延伸的长度如何均可从细胞核输出,但3'端延伸超过约10个核苷酸的前体U1 RNA在细胞质中是较差的剪接底物。3'端的结构对于RNA随后运回细胞核至关重要。这一要求确保了截短的和未完全加工的U1 RNA被排除在细胞核之外。