The ribosome is a translational apparatus that comprises about 80 ribosomal proteins and four rRNAs. Recent studies reported that ribosome ubiquitination is crucial for translational regulation and ribosome-associated quality control (RQC). However, little is known about the dynamics of ribosome ubiquitination under complex biological processes of multicellular organisms. To explore ribosome ubiquitination during animal development, we generated a zebrafish strain that expresses a FLAG-tagged ribosomal protein Rpl36/eL36 from its endogenous locus. We examined ribosome ubiquitination during zebrafish development by combining affinity purification of ribosomes from rpl36-FLAG zebrafish embryos with immunoblotting analysis. Our findings showed that the ubiquitination of ribosomal proteins dynamically changed as development proceeded. We also showed that during zebrafish development, the ribosome was ubiquitinated by Znf598, an E3 ubiquitin ligase that activates RQC. Ribosomal protein Rps10/eS10 was found to be a key ubiquitinated protein during development. Furthermore, we showed that Rps10/eS10 ubiquitination-site mutations reduced the overall ubiquitination pattern of the ribosome. These results demonstrate the complexity and dynamics of ribosome ubiquitination during zebrafish development.

中文翻译：

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Znf598介导的Rps10/eS10泛素化有助于斑马鱼发育过程中核糖体泛素化动态

核糖体是一种翻译装置，包含约 80 种核糖体蛋白和 4 种 rRNA。最近的研究表明，核糖体泛素化对于翻译调控和核糖体相关质量控制（RQC）至关重要。然而，人们对多细胞生物复杂生物过程中核糖体泛素化的动态知之甚少。为了探索动物发育过程中核糖体泛素化，我们培育了一种斑马鱼品系，该品系从其内源基因座表达 FLAG 标记的核糖体蛋白 Rpl36/eL36。我们通过将rpl36 -FLAG 斑马鱼胚胎核糖体的亲和纯化与免疫印迹分析相结合，检查了斑马鱼发育过程中的核糖体泛素化。我们的研究结果表明，核糖体蛋白的泛素化随着发育的进行而动态变化。我们还表明，在斑马鱼发育过程中，核糖体被 Znf598 泛素化，Znf598 是一种激活 RQC 的 E3 泛素连接酶。核糖体蛋白 Rps10/eS10 被发现是发育过程中关键的泛素化蛋白。此外，我们发现 Rps10/eS10 泛素化位点突变降低了核糖体的整体泛素化模式。这些结果证明了斑马鱼发育过程中核糖体泛素化的复杂性和动态性。