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Identification of small ORFs in vertebrates using ribosome footprinting and evolutionary conservation. In these disparate systems, both studies identified many new examples of translational control, uORF translation and the translation of many sORFs and alternative ORFs in genomic regions that were thought to be non-coding.īazzini, A. References 11 and 12 describe the application of ribosome profiling to physiological dynamic cellular processes, the HCMV infection cycle in human cells and meiosis in budding yeast, respectively. High-resolution view of the yeast meiotic program revealed by ribosome profiling. Ribosome profiling of mouse embryonic stem cells reveals the complexity and dynamics of mammalian proteomes. Polypeptide chain initiation: nucleotide sequences of the three ribosomal binding sites in bacteriophage R17 RNA. Ribosome pausing and stacking during translation of a eukaryotic mRNA. This work defines the ribosome profiling method and details its specificity, precision and utility. Genome-wide analysis in vivo of translation with nucleotide resolution using ribosome profiling. Ribosome-mediated specificity in Hox mRNA translation and vertebrate tissue patterning. Ribosomal protein SA haploinsufficiency in humans with isolated congenital asplenia. Ribosome biogenesis in skeletal development and the pathogenesis of skeletal disorders. Nucleolar stress in Diamond Blackfan anemia pathophysiology. Repeat-associated non-ATG (RAN) translation in neurological disease. These include novel short peptides and alternative isoforms of characterized proteins, the vast majority of which are currently of unknown function. Ribosome profiling is the first tool available for the experimental annotation of translated ORFs and has enabled the discovery of a wide range of new translation products. Proximity-specific ribosome profiling is based on localized labelling of ribosome populations within cells and enables in vivo measurement of translation at specific organelles or subcellular structures. Ribosome profiling enables instantaneous rather than steady-state measurement and is thus a particularly valuable tool for the study of gene expression over dynamic processes. The method provides quantification of levels of new protein synthesis, as well as information about ribosome positions that can be used to infer details about translation mechanism or to identify translated open reading frames (ORFs). Ribosome profiling is a deep-sequencing-based tool that allows the detailed measurement of translation globally and in vivo.