Ribosome profiling and RNA-Sequencing of wild type and Pelota knockout mouse epidermis

mRNA quality control mechanisms ensure fidelity of protein translation. An evolutionarily conserved component of the quality control machinery, Dom34/Pelota (Pelo), rescues stalled ribosomes. Here we show that Pelo is required for mammalian epidermal homeostasis. Conditional deletion in murine epidermal stem cells expressing Lrig1 results in hyperproliferation and abnormal differentiation, whereas deletion in other stem cells does not. Loss of Pelo results in truncated ribosome footprints and global upregulation of translation. Translational inhibition by rapamycin-mediated down regulation of mTOR rescues the epidermal phenotype. Our study reveals a novel role for the ribosome-rescue machinery in mammalian tissue homeostasis and an unanticipated specificity in its impact on different stem cell populations. Analysis of ribosome profiling and RNA sequencing data from primary mouse epidermis; manipulation by transgenic knockout of Pelota (PELO).mRNA质量控制机制可保障蛋白质翻译的保真度。作为质量控制系统中进化保守的组分，Dom34/Pelota（Pelo）能够解救停滞的核糖体。本研究证实，Pelo对哺乳动物表皮稳态不可或缺。在表达Lrig1的小鼠表皮干细胞中条件性敲除Pelo，会引发细胞过度增殖与分化异常，而在其他干细胞群体中进行该敲除则无此类表型。Pelo缺失会导致核糖体足迹片段截短，并引发全局翻译水平上调。通过雷帕霉素介导的mTOR通路下调以实现翻译抑制，可挽救该表皮表型。本研究揭示了核糖体解救系统在哺乳动物组织稳态中的全新功能，以及其对不同干细胞群体影响的未预期特异性。本研究对原代小鼠表皮的核糖体谱分析（ribosome profiling）与RNA测序数据展开了分析，并通过转基因敲除Pelota（PELO）开展了功能干预。