Goldfeder Mauricio B, Oliveira Carla C
Department of Biochemistry, Chemistry Institute, University of São Paulo, 748 Av. Prof. Lineu Prestes, São Paulo, SP, Brazil.
J Biol Chem. 2008 Feb 1;283(5):2644-53. doi: 10.1074/jbc.M707885200. Epub 2007 Nov 1.
U3 snoRNA is transcribed from two intron-containing genes in yeast, snR17A and snR17B. Although the assembly of the U3 snoRNP has not been precisely determined, at least some of the core box C/D proteins are known to bind pre-U3 co-transcriptionally, thereby affecting splicing and 3'-end processing of this snoRNA. We identified the interaction between the box C/D assembly factor Nop17p and Cwc24p, a novel yeast RING finger protein that had been previously isolated in a complex with the splicing factor Cef1p. Here we show that, consistent with the protein interaction data, Cwc24p localizes to the cell nucleus, and its depletion leads to the accumulation of both U3 pre-snoRNAs. U3 snoRNA is involved in the early cleavages of 35 S pre-rRNA, and the defective splicing of pre-U3 detected in cells depleted of Cwc24p causes the accumulation of the 35 S precursor rRNA. These results led us to the conclusion that Cwc24p is involved in pre-U3 snoRNA splicing, indirectly affecting pre-rRNA processing.
U3小核仁RNA(snoRNA)由酵母中两个含内含子的基因snR17A和snR17B转录而来。尽管U3小核仁核糖核蛋白颗粒(snoRNP)的组装尚未精确确定,但已知至少一些核心盒C/D蛋白在转录过程中与前体U3结合,从而影响该snoRNA的剪接和3'端加工。我们鉴定了盒C/D组装因子Nop17p与Cwc24p之间的相互作用,Cwc24p是一种新型酵母环状结构域蛋白,先前已在与剪接因子Cef1p的复合物中分离出来。在这里我们表明,与蛋白质相互作用数据一致,Cwc24p定位于细胞核,其缺失导致两种U3前体snoRNA的积累。U3 snoRNA参与35S前体核糖体RNA(pre-rRNA)的早期切割,在缺失Cwc24p的细胞中检测到的前体U3剪接缺陷导致35S前体rRNA的积累。这些结果使我们得出结论,Cwc24p参与前体U3 snoRNA的剪接,间接影响前体rRNA的加工。