Interactions between the mRNA and Rps3/uS3 at the entry tunnel of the ribosomal small subunit are important for no-go decay
| Autor: | Kyusik Q. Kim, Jessica K. Qiu, Carrie L. Simms, Liewei L. Yan, Hani S. Zaher |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Models Molecular Cancer Research Protein Conformation RNA Stability Peptide Chain Elongation Translational Cell Cycle Proteins QH426-470 Ribosome Biochemistry Protein structure Post-Translational Modification Genetics (clinical) Gel Electrophoresis Northern Analysis Messenger RNA Fungal genetics Eukaryota Cell biology Ribosome Subunits Small Nucleic acids Deletion Mutation Cellular Structures and Organelles Research Article Ribosomal Proteins Saccharomyces cerevisiae Proteins Genes Fungal Molecular Probe Techniques Saccharomyces cerevisiae Biology Research and Analysis Methods 03 medical and health sciences Electrophoretic Techniques Ribosomal protein GTP-Binding Proteins Polysome Endoribonucleases Genetics Amino Acid Sequence RNA Messenger Molecular Biology Techniques Molecular Biology Ecology Evolution Behavior and Systematics Adaptor Proteins Signal Transducing Sequence Homology Amino Acid Ubiquitination Organisms Fungi Biology and Life Sciences Proteins RNA Fungal Cell Biology Ribosomal RNA Yeast 030104 developmental biology Amino Acid Substitution Ribosome Subunits Polyribosomes Mutation Mutagenesis Site-Directed RNA Ribosomes |
| Zdroj: | PLoS Genetics PLoS Genetics, Vol 14, Iss 11, p e1007818 (2018) |
| ISSN: | 1553-7404 |
| Popis: | No-go Decay (NGD) is a process that has evolved to deal with stalled ribosomes resulting from structural blocks or aberrant mRNAs. The process is distinguished by an endonucleolytic cleavage prior to degradation of the transcript. While many of the details of the pathway have been described, the identity of the endonuclease remains unknown. Here we identify residues of the small subunit ribosomal protein Rps3 that are important for NGD by affecting the cleavage reaction. Mutation of residues within the ribosomal entry tunnel that contact the incoming mRNA leads to significantly reduced accumulation of cleavage products, independent of the type of stall sequence, and renders cells sensitive to damaging agents thought to trigger NGD. These phenotypes are distinct from those seen in combination with other NGD factors, suggesting a separate role for Rps3 in NGD. Conversely, ribosomal proteins ubiquitination is not affected by rps3 mutations, indicating that upstream ribosome quality control (RQC) events are not dependent on these residues. Together, these results suggest that Rps3 is important for quality control on the ribosome and strongly supports the notion that the ribosome itself plays a central role in the endonucleolytic cleavage reaction during NGD. Author summary In all organisms, optimum cellular fitness depends on the ability of cells to recognize and degrade aberrant molecules. Messenger RNA is subject to alterations and, as a result, often presents roadblocks for the translating ribosomes. It is not surprising, then, that organisms evolved pathways to resolve these valuable stuck ribosomes. In eukaryotes, this process is called no-go decay (NGD) because it is coupled with decay of mRNAs that are associated with ribosomes that do not ‘go’. This decay process initiates with cleavage of the mRNA near the stall site, but some important details about this reaction are lacking. Here, we show that the ribosome itself is very central to the cleavage reaction. In particular, we identified a pair of residues of a ribosomal protein to be important for cleavage efficiency. These observations are consistent with prior structural studies showing that the residues make intimate contacts with the incoming mRNA in the entry tunnel. Altogether our data provide important clues about this quality-control pathway and suggest that the endonuclease not only recognizes stalled ribosomes but may have coevolved with the translation machinery to take advantage of certain residues of the ribosome to fulfill its function. |
| Databáze: | OpenAIRE |
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