Kemmler Stefan, Occhipinti Laura, Veisu Maria, Panse Vikram Govind
Institute of Biochemistry, ETH Zürich, CH-8093 Zürich, Switzerland.
J Cell Biol. 2009 Sep 21;186(6):863-80. doi: 10.1083/jcb.200904111.
Before entering translation, preribosomal particles undergo sequential late maturation steps. In the case of pre-60S particles, these steps involve the release of shuttling maturation factors and transport receptors. In this study, we report a new maturation step in the 60S biogenesis pathway in budding yeast. We show that efficient release of the nucleolar/nuclear ribosomal-like protein Mrt4 (homologous to the acidic ribosomal P-protein Rpp0) from pre-60S particles requires the highly conserved protein Yvh1, which associates only with late pre-60S particles. Cell biological and biochemical analyses reveal that Mrt4 fails to dissociate from late pre-60S particles in yvh1Delta cells, inducing a delay in nuclear pre-ribosomal RNA processing and a pre-60S export defect in yvh1Delta cells. Moreover, we have isolated gain of function alleles of Mrt4 that specifically bypass the requirement for Yvh1 and rescue all yvh1Delta-associated phenotypes. Together, our data suggest that Yvh1-mediated release of Mrt4 precedes cytoplasmic loading of Rpp0 on pre-60S particles and is an obligatory late step toward construction of translation-competent 60S subunits.
在进入翻译过程之前,前核糖体颗粒会经历一系列后期成熟步骤。对于前60S颗粒而言,这些步骤涉及穿梭成熟因子和转运受体的释放。在本研究中,我们报道了出芽酵母60S生物合成途径中的一个新的成熟步骤。我们发现,从前60S颗粒中高效释放核仁/核核糖体样蛋白Mrt4(与酸性核糖体P蛋白Rpp0同源)需要高度保守的蛋白Yvh1,而Yvh1仅与后期前60S颗粒相关联。细胞生物学和生化分析表明,在yvh1Δ细胞中,Mrt4无法从后期前60S颗粒上解离,导致核前核糖体RNA加工延迟以及yvh1Δ细胞中前60S的输出缺陷。此外,我们分离出了Mrt4的功能获得性等位基因,这些等位基因可特异性绕过对Yvh1的需求并挽救所有与yvh1Δ相关的表型。总之,我们的数据表明,Yvh1介导的Mrt4释放先于Rpp0在前60S颗粒上的细胞质加载,并且是构建具有翻译能力的60S亚基的一个必不可少的后期步骤。