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Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA

The bacterial enzyme tRNA 2-selenouridine synthase (SelU) is responsible for the conversion of 5-substituted 2-thiouridine (R5S2U), present in the anticodon of some bacterial tRNAs, into 5-substituted 2-selenouridine (R5Se2U). We have already demonstrated using synthetic RNAs that transformation S2U...

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Autores principales: Szczupak, Patrycja, Sierant, Malgorzata, Wielgus, Ewelina, Radzikowska-Cieciura, Ewa, Kulik, Katarzyna, Krakowiak, Agnieszka, Kuwerska, Paulina, Leszczynska, Grazyna, Nawrot, Barbara
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2022
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105526/
https://www.ncbi.nlm.nih.gov/pubmed/35563829
http://dx.doi.org/10.3390/cells11091522
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author Szczupak, Patrycja
Sierant, Malgorzata
Wielgus, Ewelina
Radzikowska-Cieciura, Ewa
Kulik, Katarzyna
Krakowiak, Agnieszka
Kuwerska, Paulina
Leszczynska, Grazyna
Nawrot, Barbara
author_facet Szczupak, Patrycja
Sierant, Malgorzata
Wielgus, Ewelina
Radzikowska-Cieciura, Ewa
Kulik, Katarzyna
Krakowiak, Agnieszka
Kuwerska, Paulina
Leszczynska, Grazyna
Nawrot, Barbara
author_sort Szczupak, Patrycja
collection PubMed
description The bacterial enzyme tRNA 2-selenouridine synthase (SelU) is responsible for the conversion of 5-substituted 2-thiouridine (R5S2U), present in the anticodon of some bacterial tRNAs, into 5-substituted 2-selenouridine (R5Se2U). We have already demonstrated using synthetic RNAs that transformation S2U→Se2U is a two-step process, in which the S2U-RNA is geranylated and the resulting geS2U-RNA is selenated. Currently, the question is how SelU recognizes its substrates and what the cellular pathway of R5S2U→R5Se2U conversion is in natural tRNA. In the study presented here, we characterized the SelU substrate requirements, identified SelU-associated tRNAs and their specific modifications in the wobble position. Finally, we explained the sequence of steps in the selenation of tRNA. The S2U position within the RNA chain, the flanking sequence of the modification, and the length of the RNA substrate, all have a key influence on the recognition by SelU. MST data on the affinity of SelU to individual RNAs confirmed the presumed process. SelU binds the R5S2U-tRNA and then catalyzes its geranylation to the R5geS2U-tRNA, which remains bound to the enzyme and is selenated in the next step of the transformation. Finally, the R5Se2U-tRNA leaves the enzyme and participates in the translation process. The enzyme does not directly catalyze the R5S2U-tRNA selenation and the R5geS2U-tRNA is the intermediate product in the linear sequence of reactions.
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spelling pubmed-91055262022-05-14 Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA Szczupak, Patrycja Sierant, Malgorzata Wielgus, Ewelina Radzikowska-Cieciura, Ewa Kulik, Katarzyna Krakowiak, Agnieszka Kuwerska, Paulina Leszczynska, Grazyna Nawrot, Barbara Cells Article The bacterial enzyme tRNA 2-selenouridine synthase (SelU) is responsible for the conversion of 5-substituted 2-thiouridine (R5S2U), present in the anticodon of some bacterial tRNAs, into 5-substituted 2-selenouridine (R5Se2U). We have already demonstrated using synthetic RNAs that transformation S2U→Se2U is a two-step process, in which the S2U-RNA is geranylated and the resulting geS2U-RNA is selenated. Currently, the question is how SelU recognizes its substrates and what the cellular pathway of R5S2U→R5Se2U conversion is in natural tRNA. In the study presented here, we characterized the SelU substrate requirements, identified SelU-associated tRNAs and their specific modifications in the wobble position. Finally, we explained the sequence of steps in the selenation of tRNA. The S2U position within the RNA chain, the flanking sequence of the modification, and the length of the RNA substrate, all have a key influence on the recognition by SelU. MST data on the affinity of SelU to individual RNAs confirmed the presumed process. SelU binds the R5S2U-tRNA and then catalyzes its geranylation to the R5geS2U-tRNA, which remains bound to the enzyme and is selenated in the next step of the transformation. Finally, the R5Se2U-tRNA leaves the enzyme and participates in the translation process. The enzyme does not directly catalyze the R5S2U-tRNA selenation and the R5geS2U-tRNA is the intermediate product in the linear sequence of reactions. MDPI 2022-05-02 /pmc/articles/PMC9105526/ /pubmed/35563829 http://dx.doi.org/10.3390/cells11091522 Text en © 2022 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Szczupak, Patrycja
Sierant, Malgorzata
Wielgus, Ewelina
Radzikowska-Cieciura, Ewa
Kulik, Katarzyna
Krakowiak, Agnieszka
Kuwerska, Paulina
Leszczynska, Grazyna
Nawrot, Barbara
Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA
title Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA
title_full Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA
title_fullStr Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA
title_full_unstemmed Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA
title_short Escherichia coli tRNA 2-Selenouridine Synthase (SelU): Elucidation of Substrate Specificity to Understand the Role of S-Geranyl-tRNA in the Conversion of 2-Thio- into 2-Selenouridines in Bacterial tRNA
title_sort escherichia coli trna 2-selenouridine synthase (selu): elucidation of substrate specificity to understand the role of s-geranyl-trna in the conversion of 2-thio- into 2-selenouridines in bacterial trna
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9105526/
https://www.ncbi.nlm.nih.gov/pubmed/35563829
http://dx.doi.org/10.3390/cells11091522
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