Faber Alex W, Van Dijk Marie, Raué Hendrik A, Vos Jan C
Department of Biochemistry and Molecular Biology, Instituut Moleculair Biologische Wetenschappen, Vrije Universiteit, Amsterdam, The Netherlands.
RNA. 2002 Sep;8(9):1095-101. doi: 10.1017/s1355838202021027.
Saccharomyces cerevisiae contains three nonessential genes (NGL1, NGL2, and NGL3) that encode proteins containing a domain with similarity to a Mg(2+)-dependent endonuclease motif present in the mRNA deadenylase Ccr4p. We have investigated a possible role of these proteins in rRNA processing, because for many of the pre-rRNA processing steps, the identity of the responsible nuclease remains elusive. Analysis of RNA isolated from cells in which the NGL2 gene has been inactivated (ngl2delta) demonstrates that correct 3'-end formation of 5.8S rRNA at site E is strictly dependent on Ngl2p. No role in pre-rRNA processing could be assigned to Ngl1p and Ngl3p. The 3'-extended 5.8S rRNA formed in the ngl2delta mutant is slightly shorter than the 6S precursor previously shown to accumulate upon combined deletion of the 3' --> 5' exonuclease-encoding REX1 and REX2 genes or upon depletion of the exosomal subunits Rrp40p or Rrp45p. Thus, our data add a further component to the set of nucleases required for correct 3'-end formation of yeast 5.8S rRNA.
酿酒酵母包含三个非必需基因(NGL1、NGL2和NGL3),这些基因编码的蛋白质含有一个与mRNA脱腺苷酶Ccr4p中存在的Mg(2+)依赖性核酸内切酶基序相似的结构域。我们研究了这些蛋白质在rRNA加工中的可能作用,因为对于许多前体rRNA加工步骤,负责的核酸酶的身份仍然难以捉摸。对从NGL2基因已失活(ngl2δ)的细胞中分离的RNA进行分析表明,5.8S rRNA在E位点的正确3'末端形成严格依赖于Ngl2p。Ngl1p和Ngl3p在前体rRNA加工中没有作用。在ngl2δ突变体中形成的3'延伸的5.8S rRNA比先前显示在3'→5'外切核酸酶编码基因REX1和REX2联合缺失或外体亚基Rrp40p或Rrp45p耗尽时积累的6S前体稍短。因此,我们的数据为酵母5.8S rRNA正确3'末端形成所需的核酸酶集合增加了另一个组成部分。