The Effect of tRNA
| Autor: | Tsutomu Suzuki, Uschi Reuter, Noelia Fradejas-Villar, Wenchao Zhao, Kenjyo Miyauchi, Rainer Knoll, Robert McFarland, Mark Helm, Simon Bohleber, Robert W. Taylor, Yuriko Sakaguchi, Annika Kotter, Ulrich Schweizer, Yufeng Mo |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
GPX1
medicine.disease_cause law.invention tRNA[Ser]Sec Mice RNA Transfer law Biology (General) Trit1 Selenoproteins Spectroscopy chemistry.chemical_classification Neurons Mutation Chemistry Translation (biology) General Medicine Computer Science Applications Blot Liver Transfer RNA Recombinant DNA QH301-705.5 isopentenylation Catalysis Article Cell Line Inorganic Chemistry Selenium Selenoprotein P medicine Animals Humans Cysteine Physical and Theoretical Chemistry Molecular Biology QD1-999 Alkyl and Aryl Transferases Organic Chemistry Phosphotransferases Molecular biology In vitro Selenocysteine Protein Biosynthesis Hepatocytes Selenoprotein Ribosomes |
| Zdroj: | International Journal of Molecular Sciences Volume 22 Issue 21 International Journal of Molecular Sciences, Vol 22, Iss 11454, p 11454 (2021) |
| ISSN: | 1422-0067 |
| Popis: | Transfer RNA[Ser]Sec carries multiple post-transcriptional modifications. The A37G mutation in tRNA[Ser]Sec abrogates isopentenylation of base 37 and has a profound effect on selenoprotein expression in mice. Patients with a homozygous pathogenic p.R323Q variant in tRNA-isopentenyl-transferase (TRIT1) show a severe neurological disorder, and hence we wondered whether selenoprotein expression was impaired. Patient fibroblasts with the homozygous p.R323Q variant did not show a general decrease in selenoprotein expression. However, recombinant human TRIT1R323Q had significantly diminished activities towards several tRNA substrates in vitro. We thus engineered mice conditionally deficient in Trit1 in hepatocytes and neurons. Mass-spectrometry revealed that hypermodification of U34 to mcm5Um occurs independently of isopentenylation of A37 in tRNA[Ser]Sec. Western blotting and 75Se metabolic labeling showed only moderate effects on selenoprotein levels and 75Se incorporation. A detailed analysis of Trit1-deficient liver using ribosomal profiling demonstrated that UGA/Sec re-coding was moderately affected in Selenop, Txnrd1, and Sephs2, but not in Gpx1. 2′O-methylation of U34 in tRNA[Ser]Sec depends on FTSJ1, but does not affect UGA/Sec re-coding in selenoprotein translation. Taken together, our results show that a lack of isopentenylation of tRNA[Ser]Sec affects UGA/Sec read-through but differs from a A37G mutation. |
| Databáze: | OpenAIRE |
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