Walther T N, Wittop Koning T H, Schümperli D, Müller B
Abteilung für Entwicklungsbiologie, Zoologisches Institut, Universität Bern, Switzerland.
RNA. 1998 Sep;4(9):1034-46. doi: 10.1017/s1355838298971771.
Histone RNA 3' processing in vitro produces one or more 5' cleavage products corresponding to the mature histone mRNA 3' end, and a group of 3' cleavage products whose 5' ends are mostly located several nucleotides downstream of the mRNA 3' end. The formation of these 3' products is coupled to the formation of 5' products and dependent on the U7 snRNP and a heat-labile processing factor. These short 3' products therefore are a true and general feature of the processing reaction. Identical 3' products are also formed from a model RNA containing all spacer nucleotides downstream of the mature mRNA 3' end, but no sequences from the mature mRNA. Again, this reaction is dependent on both the U7 snRNP and a heat-labile factor. Unlike the processing with a full-length histone pre-mRNA, this reaction produces only 3' but no 5' fragments. In addition, product formation is inhibited by addition of cap structures at the model RNA 5' end, indicating that product formation occurs by 5'-3' exonucleolytic degradation. This degradation of a model 3' product by a 5'-3' exonuclease suggests a mechanism for the release of the U7 snRNP after processing by shortening the cut-off histone spacer sequences base paired to U7 RNA.
体外组蛋白RNA 3'加工产生一个或多个对应于成熟组蛋白mRNA 3'末端的5'切割产物,以及一组3'切割产物,其5'末端大多位于mRNA 3'末端下游的几个核苷酸处。这些3'产物的形成与5'产物的形成偶联,并依赖于U7 snRNP和一个热不稳定加工因子。因此,这些短的3'产物是加工反应的一个真实且普遍的特征。从一个包含成熟mRNA 3'末端下游所有间隔核苷酸但没有成熟mRNA序列的模型RNA中也能形成相同的3'产物。同样,这个反应依赖于U7 snRNP和一个热不稳定因子。与全长组蛋白前体mRNA的加工不同,这个反应只产生3'片段而不产生5'片段。此外,在模型RNA 5'末端添加帽结构会抑制产物形成,这表明产物形成是通过5'-3'核酸外切酶降解发生的。5'-3'核酸外切酶对模型3'产物的这种降解提示了一种机制,即在加工后通过缩短与U7 RNA碱基配对的截断组蛋白间隔序列来释放U7 snRNP。