論文 - 詳細
| RRC ID | 69978 |
|---|---|
| 著者 | Kimura Y, Saito H, Osaki T, Ikegami Y, Wakigawa T, Ikeuchi Y, Iwasaki S. |
| タイトル | Mito-FUNCAT-FACS reveals cellular heterogeneity in mitochondrial translation. |
| ジャーナル | RNA |
| Abstract |
Mitochondria possess their own genome that encodes components of oxidative phosphorylation (OXPHOS) complexes, and mitochondrial ribosomes within the organelle translate the mRNAs expressed from the mitochondrial genome. Given the differential OXPHOS activity observed in diverse cell types, cell growth conditions, and other circumstances, cellular heterogeneity in mitochondrial translation can be expected. Although individual protein products translated in mitochondria have been monitored, the lack of techniques that address the variation in overall mitochondrial protein synthesis in cell populations poses analytic challenges. Here, we adapted mitochondrial-specific fluorescent noncanonical amino acid tagging (FUNCAT) for use with fluorescence-activated cell sorting (FACS) and developed mito-FUNCAT-FACS. The click chemistry-compatible methionine analog L-homopropargylglycine (HPG) enabled the metabolic labeling of newly synthesized proteins. In the presence of cytosolic translation inhibitors, HPG was selectively incorporated into mitochondrial nascent proteins and conjugated to fluorophores via the click reaction (mito-FUNCAT). The application of in situ mito-FUNCAT to flow cytometry allowed us to separate changes in net mitochondrial translation activity from those of the organelle mass and detect variations in mitochondrial translation in cancer cells. Our approach provides a useful methodology for examining mitochondrial protein synthesis in individual cells. |
| 巻・号 | 28(6) |
| ページ | 895-904 |
| 公開日 | 2022-6-1 |
| DOI | 10.1261/rna.079097.122 |
| PII | rna.079097.122 |
| PMID | 35256452 |
| PMC | PMC9074903 |
| MeSH | Amino Acids* / chemistry Flow Cytometry Mitochondria / genetics Mitochondria / metabolism Mitochondrial Proteins / genetics Mitochondrial Proteins / metabolism Protein Biosynthesis* |
| IF | 4.32 |
| オルトメトリクス指標 |
オルトメトリクス指標項目
|
| 最多言及媒体 | X(Twitter) |
| 各媒体での言及数の合計 | 44 |
| 過去6か月間でのオルトメトリクス指標の変動値 | 0.0 |
| リソース情報 | |
| ヒト・動物細胞 | HeLa.S3(RCB1525) |