Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coli
Lamotrigine (Ltg), an anticonvulsant drug, targets initiation factor 2 (IF2), compromises ribosome biogenesis and causes toxicity to Escherichia coli. However, our understanding of Ltg toxicity in E. coli remains unclear. While our in vitro assays reveal no effects of Ltg on the ribosome-dependent G...
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| Format: | Article |
| Language: | English |
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Taylor & Francis Group
2023-12-01
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| Series: | RNA Biology |
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| Online Access: | http://dx.doi.org/10.1080/15476286.2023.2253395 |
| _version_ | 1797403963896627200 |
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| author | Sudhir Singh Kuldeep Lahry Chandra Sekhar Mandava Jitendra Singh Riyaz Ahmad Shah Suparna Sanyal Umesh Varshney |
| author_facet | Sudhir Singh Kuldeep Lahry Chandra Sekhar Mandava Jitendra Singh Riyaz Ahmad Shah Suparna Sanyal Umesh Varshney |
| author_sort | Sudhir Singh |
| collection | DOAJ |
| description | Lamotrigine (Ltg), an anticonvulsant drug, targets initiation factor 2 (IF2), compromises ribosome biogenesis and causes toxicity to Escherichia coli. However, our understanding of Ltg toxicity in E. coli remains unclear. While our in vitro assays reveal no effects of Ltg on the ribosome-dependent GTPase activity of IF2 or its role in initiation as measured by dipeptide formation in a fast kinetics assay, the in vivo experiments show that Ltg causes accumulation of the 17S precursor of 16S rRNA and leads to a decrease in polysome levels in E. coli. IF2 overexpression in E. coli increases Ltg toxicity. However, the overexpression of initiator tRNA (i-tRNA) protects it from the Ltg toxicity. The depletion of i-tRNA or overexpression of its 3GC mutant (lacking the characteristic 3GC base pairs in anticodon stem) enhances Ltg toxicity, and this enhancement in toxicity is synthetic with IF2 overexpression. The Ltg treatment itself causes a detectable increase in IF2 levels in E. coli and allows initiation with an elongator tRNA, suggesting compromise in the fidelity/specificity of IF2 function. Also, Ltg causes increased accumulation of ribosome-binding factor A (RbfA) on 30S ribosomal subunit. Based on our genetic and biochemical investigations, we show that Ltg compromises the function of i-tRNA/IF2 complex in ribosome maturation. |
| first_indexed | 2024-03-09T02:46:13Z |
| format | Article |
| id | doaj.art-56e200cc0fe04e10af7d172e7a9270b7 |
| institution | Directory Open Access Journal |
| issn | 1547-6286 1555-8584 |
| language | English |
| last_indexed | 2024-03-09T02:46:13Z |
| publishDate | 2023-12-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | RNA Biology |
| spelling | doaj.art-56e200cc0fe04e10af7d172e7a9270b72023-12-05T16:09:52ZengTaylor & Francis GroupRNA Biology1547-62861555-85842023-12-0120168169210.1080/15476286.2023.22533952253395Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coliSudhir Singh0Kuldeep Lahry1Chandra Sekhar Mandava2Jitendra Singh3Riyaz Ahmad Shah4Suparna Sanyal5Umesh Varshney6Indian Institute of ScienceIndian Institute of ScienceUppsala UniversityIndian Institute of ScienceIndian Institute of ScienceUppsala UniversityIndian Institute of ScienceLamotrigine (Ltg), an anticonvulsant drug, targets initiation factor 2 (IF2), compromises ribosome biogenesis and causes toxicity to Escherichia coli. However, our understanding of Ltg toxicity in E. coli remains unclear. While our in vitro assays reveal no effects of Ltg on the ribosome-dependent GTPase activity of IF2 or its role in initiation as measured by dipeptide formation in a fast kinetics assay, the in vivo experiments show that Ltg causes accumulation of the 17S precursor of 16S rRNA and leads to a decrease in polysome levels in E. coli. IF2 overexpression in E. coli increases Ltg toxicity. However, the overexpression of initiator tRNA (i-tRNA) protects it from the Ltg toxicity. The depletion of i-tRNA or overexpression of its 3GC mutant (lacking the characteristic 3GC base pairs in anticodon stem) enhances Ltg toxicity, and this enhancement in toxicity is synthetic with IF2 overexpression. The Ltg treatment itself causes a detectable increase in IF2 levels in E. coli and allows initiation with an elongator tRNA, suggesting compromise in the fidelity/specificity of IF2 function. Also, Ltg causes increased accumulation of ribosome-binding factor A (RbfA) on 30S ribosomal subunit. Based on our genetic and biochemical investigations, we show that Ltg compromises the function of i-tRNA/IF2 complex in ribosome maturation.http://dx.doi.org/10.1080/15476286.2023.2253395ribosomelamotrigineltginitiation factor 2initiator trna |
| spellingShingle | Sudhir Singh Kuldeep Lahry Chandra Sekhar Mandava Jitendra Singh Riyaz Ahmad Shah Suparna Sanyal Umesh Varshney Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coli RNA Biology ribosome lamotrigine ltg initiation factor 2 initiator trna |
| title | Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coli |
| title_full | Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coli |
| title_fullStr | Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coli |
| title_full_unstemmed | Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coli |
| title_short | Lamotrigine compromises the fidelity of initiator tRNA recruitment to the ribosomal P-site by IF2 and the RbfA release from 30S ribosomes in Escherichia coli |
| title_sort | lamotrigine compromises the fidelity of initiator trna recruitment to the ribosomal p site by if2 and the rbfa release from 30s ribosomes in escherichia coli |
| topic | ribosome lamotrigine ltg initiation factor 2 initiator trna |
| url | http://dx.doi.org/10.1080/15476286.2023.2253395 |
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