Perreault Audrey, Bellemer Clément, Bachand Francois
RNA Group, Department of Biochemistry, Université de Sherbrooke, Québec, Canada.
Nucleic Acids Res. 2008 Nov;36(19):6132-42. doi: 10.1093/nar/gkn625. Epub 2008 Sep 27.
Ribosome biogenesis is an evolutionarily conserved pathway that requires ribosomal and nonribosomal proteins. Here, we investigated the role of the ribosomal protein S2 (Rps2) in fission yeast ribosome synthesis. As for many budding yeast ribosomal proteins, Rps2 was essential for cell viability in fission yeast and the genetic depletion of Rps2 caused a complete inhibition of 40S ribosomal subunit production. The pattern of pre-rRNA processing upon depletion of Rps2 revealed a reduction of 27SA(2) pre-rRNAs and the concomitant production of 21S rRNA precursors, consistent with a role for Rps2 in efficient cleavage at site A(2) within the 32S pre-rRNA. Importantly, kinetics of pre-rRNA accumulation as determined by rRNA pulse-chases assays indicated that a small fraction of 35S precursors matured into 20S-containing particles, suggesting that most 40S precursors were rapidly degraded in the absence of Rps2. Analysis of steady-state RNA levels revealed that some pre-40S particles were produced in Rps2-depleted cells, but that these precursors were retained in the nucleolus. Our findings suggest a role for Rps2 in a mechanism that monitors pre-40S export competence.
核糖体生物合成是一条进化上保守的途径,需要核糖体蛋白和非核糖体蛋白。在这里,我们研究了核糖体蛋白S2(Rps2)在裂殖酵母核糖体合成中的作用。与许多芽殖酵母核糖体蛋白一样,Rps2对裂殖酵母的细胞活力至关重要,Rps2的基因缺失导致40S核糖体亚基的产生完全受到抑制。Rps2缺失时前体rRNA的加工模式显示27SA(2)前体rRNA减少,同时产生21S rRNA前体,这与Rps2在32S前体rRNA内A(2)位点的有效切割中发挥的作用一致。重要的是,通过rRNA脉冲追踪实验确定的前体rRNA积累动力学表明,一小部分35S前体成熟为含20S的颗粒,这表明在没有Rps2的情况下,大多数40S前体迅速降解。对稳态RNA水平的分析表明,在Rps2缺失的细胞中产生了一些40S前体颗粒,但这些前体保留在核仁中。我们的研究结果表明Rps2在监测40S前体输出能力的机制中发挥作用。