Identification of distinct maturation steps involved in human 40S ribosomal subunit biosynthesis
[EN]Technical problems intrinsic to the purification of preribosome intermediates have limited our understanding of ribosome biosynthesis in humans. Addressing this issue is important given the implication of this biological process in human disease. Here we report a preribosome purification and tag...
| Authors: | , , , , , |
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| Format: | article |
| Status: | Published version |
| Publication Date: | 2020 |
| Country: | España |
| Institution: | Universidad de Salamanca (USAL) |
| Repository: | GREDOS. Repositorio Institucional de la Universidad de Salamanca |
| OAI Identifier: | oai:gredos.usal.es:10366/168969 |
| Online Access: | http://hdl.handle.net/10366/168969 |
| Access Level: | Open access |
| Keyword: | Ribosome maturation human ribosome preribosome 40S ribosomal subunit pre-rRNA human nucleolus RNA Ribosomal Proteins Staining and Labeling Exoribonucleases Humans Cell Nucleolus Cell Line HCT116 Cells HeLa Cells RNA Precursors Ribosome Subunits Nuclear Proteins 2407 Biología Celular 2415 Biología Molecular 2409 Genética 2302.05 Biosíntesis precursores del ARN nucleolo celular humanos tinción y etiquetado línea celular ARN células HCT116 células HeLa subunidades ribosómicas proteínas ribosómicas exorribonucleasas proteínas nucleares |
| Summary: | [EN]Technical problems intrinsic to the purification of preribosome intermediates have limited our understanding of ribosome biosynthesis in humans. Addressing this issue is important given the implication of this biological process in human disease. Here we report a preribosome purification and tagging strategy that overcomes some of the existing technical difficulties. Using these tools, we find that the pre-40S precursors go through two distinct maturation phases inside the nucleolus and follow a regulatory step that precedes late maturation in the cytoplasm. This regulatory step entails the intertwined actions of both PARN (a metazoan-specific ribonuclease) and RRP12 (a phylogenetically conserved 40S biogenesis factor that has acquired additional functional features in higher eukaryotes). Together, these results demonstrate the usefulness of this purification method for the dissection of ribosome biogenesis in human cells. They also identify distinct maturation stages and metazoan-specific regulatory mechanisms involved in the generation of the human 40S ribosomal subunit. |
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