The Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression
Transfer RNA<sup>[Ser]Sec</sup> carries multiple post-transcriptional modifications. The A37G mutation in tRNA<sup>[Ser]Sec</sup> abrogates isopentenylation of base 37 and has a profound effect on selenoprotein expression in mice. Patients with a homozygous pathogenic p.R323Q...
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oai:doaj.org-article:9a8205bba91f4fc3874f11f8544724c32021-11-11T16:54:58ZThe Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression10.3390/ijms2221114541422-00671661-6596https://doaj.org/article/9a8205bba91f4fc3874f11f8544724c32021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11454https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Transfer RNA<sup>[Ser]Sec</sup> carries multiple post-transcriptional modifications. The A37G mutation in tRNA<sup>[Ser]Sec</sup> 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 (<i>TRIT1</i>) 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 TRIT1<sup>R323Q</sup> had significantly diminished activities towards several tRNA substrates in vitro. We thus engineered mice conditionally deficient in <i>Trit1</i> in hepatocytes and neurons. Mass-spectrometry revealed that hypermodification of U<sub>34</sub> to mcm<sup>5</sup>Um occurs independently of isopentenylation of A<sub>37</sub> in tRNA<sup>[Ser]Sec</sup>. Western blotting and <sup>75</sup>Se metabolic labeling showed only moderate effects on selenoprotein levels and <sup>75</sup>Se incorporation. A detailed analysis of <i>Trit1</i>-deficient liver using ribosomal profiling demonstrated that UGA/Sec re-coding was moderately affected in <i>Selenop</i>, <i>Txnrd1</i>, and <i>Sephs2</i>, but not in <i>Gpx1</i>. 2′O-methylation of U<sub>34</sub> in tRNA<sup>[Ser]Sec</sup> 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<sup>[Ser]Sec</sup> affects UGA/Sec read-through but differs from a A37G mutation.Noelia Fradejas-VillarSimon BohleberWenchao ZhaoUschi ReuterAnnika KotterMark HelmRainer KnollRobert McFarlandRobert W. TaylorYufeng MoKenjyo MiyauchiYuriko SakaguchiTsutomu SuzukiUlrich SchweizerMDPI AGarticle<i>Trit1</i>isopentenylationtRNA<sup>[Ser]Sec</sup>selenoproteinsBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11454, p 11454 (2021) |
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<i>Trit1</i> isopentenylation tRNA<sup>[Ser]Sec</sup> selenoproteins Biology (General) QH301-705.5 Chemistry QD1-999 |
| spellingShingle |
<i>Trit1</i> isopentenylation tRNA<sup>[Ser]Sec</sup> selenoproteins Biology (General) QH301-705.5 Chemistry QD1-999 Noelia Fradejas-Villar Simon Bohleber Wenchao Zhao Uschi Reuter Annika Kotter Mark Helm Rainer Knoll Robert McFarland Robert W. Taylor Yufeng Mo Kenjyo Miyauchi Yuriko Sakaguchi Tsutomu Suzuki Ulrich Schweizer The Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression |
| description |
Transfer RNA<sup>[Ser]Sec</sup> carries multiple post-transcriptional modifications. The A37G mutation in tRNA<sup>[Ser]Sec</sup> 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 (<i>TRIT1</i>) 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 TRIT1<sup>R323Q</sup> had significantly diminished activities towards several tRNA substrates in vitro. We thus engineered mice conditionally deficient in <i>Trit1</i> in hepatocytes and neurons. Mass-spectrometry revealed that hypermodification of U<sub>34</sub> to mcm<sup>5</sup>Um occurs independently of isopentenylation of A<sub>37</sub> in tRNA<sup>[Ser]Sec</sup>. Western blotting and <sup>75</sup>Se metabolic labeling showed only moderate effects on selenoprotein levels and <sup>75</sup>Se incorporation. A detailed analysis of <i>Trit1</i>-deficient liver using ribosomal profiling demonstrated that UGA/Sec re-coding was moderately affected in <i>Selenop</i>, <i>Txnrd1</i>, and <i>Sephs2</i>, but not in <i>Gpx1</i>. 2′O-methylation of U<sub>34</sub> in tRNA<sup>[Ser]Sec</sup> 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<sup>[Ser]Sec</sup> affects UGA/Sec read-through but differs from a A37G mutation. |
| format |
article |
| author |
Noelia Fradejas-Villar Simon Bohleber Wenchao Zhao Uschi Reuter Annika Kotter Mark Helm Rainer Knoll Robert McFarland Robert W. Taylor Yufeng Mo Kenjyo Miyauchi Yuriko Sakaguchi Tsutomu Suzuki Ulrich Schweizer |
| author_facet |
Noelia Fradejas-Villar Simon Bohleber Wenchao Zhao Uschi Reuter Annika Kotter Mark Helm Rainer Knoll Robert McFarland Robert W. Taylor Yufeng Mo Kenjyo Miyauchi Yuriko Sakaguchi Tsutomu Suzuki Ulrich Schweizer |
| author_sort |
Noelia Fradejas-Villar |
| title |
The Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression |
| title_short |
The Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression |
| title_full |
The Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression |
| title_fullStr |
The Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression |
| title_full_unstemmed |
The Effect of tRNA<sup>[Ser]Sec</sup> Isopentenylation on Selenoprotein Expression |
| title_sort |
effect of trna<sup>[ser]sec</sup> isopentenylation on selenoprotein expression |
| publisher |
MDPI AG |
| publishDate |
2021 |
| url |
https://doaj.org/article/9a8205bba91f4fc3874f11f8544724c3 |
| work_keys_str_mv |
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