Queuine Salvaging in the Human Parasite Entamoeba histolytica
Queuosine (Q) is a naturally occurring modified nucleoside that occurs in the first position of transfer RNA anticodons such as Asp, Asn, His, and Tyr. As eukaryotes lack pathways to synthesize queuine, the Q nucleobase, they must obtain it from their diet or gut microbiota. Previously, we described...
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| Format: | Article |
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Multidisciplinary Digital Publishing Institute
2022
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| Online Access: | https://hdl.handle.net/1721.1/144428 |
| _version_ | 1811079782604996608 |
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| author | Sarid, Lotem Sun, Jingjing Chittrakanwong, Jurairat Trebicz-Geffen, Meirav Ye, Jun Dedon, Peter C. Ankri, Serge |
| author2 | Massachusetts Institute of Technology. Department of Biological Engineering |
| author_facet | Massachusetts Institute of Technology. Department of Biological Engineering Sarid, Lotem Sun, Jingjing Chittrakanwong, Jurairat Trebicz-Geffen, Meirav Ye, Jun Dedon, Peter C. Ankri, Serge |
| author_sort | Sarid, Lotem |
| collection | MIT |
| description | Queuosine (Q) is a naturally occurring modified nucleoside that occurs in the first position of transfer RNA anticodons such as Asp, Asn, His, and Tyr. As eukaryotes lack pathways to synthesize queuine, the Q nucleobase, they must obtain it from their diet or gut microbiota. Previously, we described the effects of queuine on the physiology of the eukaryotic parasite <i>Entamoeba histolytica</i> and characterized the enzyme EhTGT responsible for queuine incorporation into tRNA. At present, it is unknown how <i>E. histolytica</i> salvages queuine from gut bacteria. We used liquid chromatography–mass spectrometry (LC–MS) and N-acryloyl-3-aminophenylboronic acid (APB) PAGE analysis to demonstrate that <i>E. histolytica</i> trophozoites can salvage queuine from Q or <i>E. coli</i> K12 but not from the modified <i>E. coli</i> QueC strain, which cannot produce queuine. We then examined the role of EhDUF2419, a protein with homology to DNA glycosylase, as a queuine salvage enzyme in <i>E. histolytica</i>. We found that glutathione S-transferase (GST)-EhDUF2419 catalyzed the conversion of Q into queuine. Trophozoites silenced for EhDUF2419 expression are impaired in their ability to form Q-tRNA from Q or from <i>E. coli</i>. We also observed that Q or <i>E. coli</i> K12 partially protects control trophozoites from oxidative stress (OS), but not siEhDUF2419 trophozoites. Overall, our data reveal that EhDUF2419 is central for the direct salvaging of queuine from bacteria and for the resistance of the parasite to OS. |
| first_indexed | 2024-09-23T11:20:23Z |
| format | Article |
| id | mit-1721.1/144428 |
| institution | Massachusetts Institute of Technology |
| last_indexed | 2024-09-23T11:20:23Z |
| publishDate | 2022 |
| publisher | Multidisciplinary Digital Publishing Institute |
| record_format | dspace |
| spelling | mit-1721.1/1444282023-04-11T19:51:28Z Queuine Salvaging in the Human Parasite Entamoeba histolytica Sarid, Lotem Sun, Jingjing Chittrakanwong, Jurairat Trebicz-Geffen, Meirav Ye, Jun Dedon, Peter C. Ankri, Serge Massachusetts Institute of Technology. Department of Biological Engineering Queuosine (Q) is a naturally occurring modified nucleoside that occurs in the first position of transfer RNA anticodons such as Asp, Asn, His, and Tyr. As eukaryotes lack pathways to synthesize queuine, the Q nucleobase, they must obtain it from their diet or gut microbiota. Previously, we described the effects of queuine on the physiology of the eukaryotic parasite <i>Entamoeba histolytica</i> and characterized the enzyme EhTGT responsible for queuine incorporation into tRNA. At present, it is unknown how <i>E. histolytica</i> salvages queuine from gut bacteria. We used liquid chromatography–mass spectrometry (LC–MS) and N-acryloyl-3-aminophenylboronic acid (APB) PAGE analysis to demonstrate that <i>E. histolytica</i> trophozoites can salvage queuine from Q or <i>E. coli</i> K12 but not from the modified <i>E. coli</i> QueC strain, which cannot produce queuine. We then examined the role of EhDUF2419, a protein with homology to DNA glycosylase, as a queuine salvage enzyme in <i>E. histolytica</i>. We found that glutathione S-transferase (GST)-EhDUF2419 catalyzed the conversion of Q into queuine. Trophozoites silenced for EhDUF2419 expression are impaired in their ability to form Q-tRNA from Q or from <i>E. coli</i>. We also observed that Q or <i>E. coli</i> K12 partially protects control trophozoites from oxidative stress (OS), but not siEhDUF2419 trophozoites. Overall, our data reveal that EhDUF2419 is central for the direct salvaging of queuine from bacteria and for the resistance of the parasite to OS. 2022-08-25T12:41:43Z 2022-08-25T12:41:43Z 2022-08-12 2022-08-25T11:17:37Z Article http://purl.org/eprint/type/JournalArticle https://hdl.handle.net/1721.1/144428 Cells 11 (16): 2509 (2022) PUBLISHER_CC http://dx.doi.org/10.3390/cells11162509 Creative Commons Attribution https://creativecommons.org/licenses/by/4.0/ application/pdf Multidisciplinary Digital Publishing Institute Multidisciplinary Digital Publishing Institute |
| spellingShingle | Sarid, Lotem Sun, Jingjing Chittrakanwong, Jurairat Trebicz-Geffen, Meirav Ye, Jun Dedon, Peter C. Ankri, Serge Queuine Salvaging in the Human Parasite Entamoeba histolytica |
| title | Queuine Salvaging in the Human Parasite Entamoeba histolytica |
| title_full | Queuine Salvaging in the Human Parasite Entamoeba histolytica |
| title_fullStr | Queuine Salvaging in the Human Parasite Entamoeba histolytica |
| title_full_unstemmed | Queuine Salvaging in the Human Parasite Entamoeba histolytica |
| title_short | Queuine Salvaging in the Human Parasite Entamoeba histolytica |
| title_sort | queuine salvaging in the human parasite entamoeba histolytica |
| url | https://hdl.handle.net/1721.1/144428 |
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