Ooi S L, Samarsky D A, Fournier M J, Boeke J D
Department of Molecular Biology and Genetics, Johns Hopkins University School of Medicine, Baltimore, Maryland 21205, USA.
RNA. 1998 Sep;4(9):1096-110. doi: 10.1017/s1355838298980785.
The eukaryotic small nucleolar RNAs (snoRNAs) are involved in processing of pre-rRNA and modification of rRNA nucleotides. Some snoRNAs are derived from mono- or polycistronic transcription units, whereas others are encoded in introns of protein genes. The present study addresses the role of the RNA lariat-debranching enzyme (Dbr1p) in the synthesis and function of intronic snoRNAs in the yeast Saccharomyces cerevisiae. Intronic snoRNA production was determined to depend on Dbr1p. Accumulation of mature intronic snoRNAs is reduced in a dbr1 mutant; instead, intronic snoRNAs are "trapped" within host intron lariats. Interestingly, the extent of intronic snoRNA accumulation in the form of lariats in dbr1 cells varied among different intronic snoRNAs. Intronic snoRNAs encoded within shorter introns, such as U24 and snR38, accumulate more unprocessed lariat precursors than those encoded within longer introns, e.g., U18 and snR39. This correlation was corroborated by experiments conducted with model intron:U24 snoRNA constructs. These results support a splicing-dependent exonucleolytic pathway for the biosynthesis of intronic snoRNAs. Curiously, U24 in a lariat may be functional in directing methylation of ribosomal RNA.
真核生物小核仁RNA(snoRNAs)参与前体rRNA的加工以及rRNA核苷酸的修饰。一些snoRNAs来源于单顺反子或多顺反子转录单元,而其他的则由蛋白质基因的内含子编码。本研究探讨了RNA套索脱支酶(Dbr1p)在酿酒酵母内含子snoRNAs合成及功能中的作用。研究确定内含子snoRNA的产生依赖于Dbr1p。在dbr1突变体中,成熟内含子snoRNAs的积累减少;相反,内含子snoRNAs“被困”在宿主内含子套索中。有趣的是,dbr1细胞中以套索形式存在的内含子snoRNA积累程度在不同的内含子snoRNAs之间存在差异。较短内含子(如U24和snR38)中编码的内含子snoRNAs比较长内含子(如U18和snR39)中编码的内含子snoRNAs积累更多未加工的套索前体。用模型内含子:U24 snoRNA构建体进行的实验证实了这种相关性。这些结果支持了一种依赖剪接的核酸外切酶途径用于内含子snoRNAs的生物合成。奇怪的是,套索形式的U24可能在指导核糖体RNA甲基化方面具有功能。