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RbgA ensures the correct timing in the maturation of the 50S subunits functional sites
RbgA is an essential protein for the assembly of the 50S subunit in Bacillus subtilis. Depletion of RbgA leads to the accumulation of the 45S intermediate. A strain expressing a RbgA variant with reduced GTPase activity generates spontaneous suppressor mutations in uL6. Each suppressor strain accumu...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Oxford University Press
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9638911/ https://www.ncbi.nlm.nih.gov/pubmed/35141754 http://dx.doi.org/10.1093/nar/gkac059 |
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author | Seffouh, Amal Trahan, Chirstian Wasi, Tanzila Jain, Nikhil Basu, Kaustuv Britton, Robert A Oeffinger, Marlene Ortega, Joaquin |
author_facet | Seffouh, Amal Trahan, Chirstian Wasi, Tanzila Jain, Nikhil Basu, Kaustuv Britton, Robert A Oeffinger, Marlene Ortega, Joaquin |
author_sort | Seffouh, Amal |
collection | PubMed |
description | RbgA is an essential protein for the assembly of the 50S subunit in Bacillus subtilis. Depletion of RbgA leads to the accumulation of the 45S intermediate. A strain expressing a RbgA variant with reduced GTPase activity generates spontaneous suppressor mutations in uL6. Each suppressor strain accumulates a unique 44S intermediate. We reasoned that characterizing the structure of these mutant 44S intermediates may explain why RbgA is required to catalyze the folding of the 50S functional sites. We found that in the 44S particles, rRNA helices H42 and H97, near the binding site of uL6, adopt a flexible conformation and allow the central protuberance and functional sites in the mutant 44S particles to mature in any order. Instead, the wild-type 45S particles exhibit a stable H42-H97 interaction and their functional sites always mature last. The dependence on RbgA was also less pronounced in the 44S particles. We concluded that the binding of uL6 pauses the maturation of the functional sites, but the central protuberance continues to fold. RbgA exclusively binds intermediates with a formed central protuberance and licenses the folding of the functional sites. Through this mechanism, RbgA ensures that the functional sites of the 50S mature last. |
format | Online Article Text |
id | pubmed-9638911 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Oxford University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-96389112022-11-07 RbgA ensures the correct timing in the maturation of the 50S subunits functional sites Seffouh, Amal Trahan, Chirstian Wasi, Tanzila Jain, Nikhil Basu, Kaustuv Britton, Robert A Oeffinger, Marlene Ortega, Joaquin Nucleic Acids Res NAR Breakthrough Article RbgA is an essential protein for the assembly of the 50S subunit in Bacillus subtilis. Depletion of RbgA leads to the accumulation of the 45S intermediate. A strain expressing a RbgA variant with reduced GTPase activity generates spontaneous suppressor mutations in uL6. Each suppressor strain accumulates a unique 44S intermediate. We reasoned that characterizing the structure of these mutant 44S intermediates may explain why RbgA is required to catalyze the folding of the 50S functional sites. We found that in the 44S particles, rRNA helices H42 and H97, near the binding site of uL6, adopt a flexible conformation and allow the central protuberance and functional sites in the mutant 44S particles to mature in any order. Instead, the wild-type 45S particles exhibit a stable H42-H97 interaction and their functional sites always mature last. The dependence on RbgA was also less pronounced in the 44S particles. We concluded that the binding of uL6 pauses the maturation of the functional sites, but the central protuberance continues to fold. RbgA exclusively binds intermediates with a formed central protuberance and licenses the folding of the functional sites. Through this mechanism, RbgA ensures that the functional sites of the 50S mature last. Oxford University Press 2022-02-10 /pmc/articles/PMC9638911/ /pubmed/35141754 http://dx.doi.org/10.1093/nar/gkac059 Text en © The Author(s) 2022. Published by Oxford University Press on behalf of Nucleic Acids Research. https://creativecommons.org/licenses/by-nc/4.0/This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial License (https://creativecommons.org/licenses/by-nc/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited. For commercial re-use, please contact journals.permissions@oup.com |
spellingShingle | NAR Breakthrough Article Seffouh, Amal Trahan, Chirstian Wasi, Tanzila Jain, Nikhil Basu, Kaustuv Britton, Robert A Oeffinger, Marlene Ortega, Joaquin RbgA ensures the correct timing in the maturation of the 50S subunits functional sites |
title | RbgA ensures the correct timing in the maturation of the 50S subunits functional sites |
title_full | RbgA ensures the correct timing in the maturation of the 50S subunits functional sites |
title_fullStr | RbgA ensures the correct timing in the maturation of the 50S subunits functional sites |
title_full_unstemmed | RbgA ensures the correct timing in the maturation of the 50S subunits functional sites |
title_short | RbgA ensures the correct timing in the maturation of the 50S subunits functional sites |
title_sort | rbga ensures the correct timing in the maturation of the 50s subunits functional sites |
topic | NAR Breakthrough Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9638911/ https://www.ncbi.nlm.nih.gov/pubmed/35141754 http://dx.doi.org/10.1093/nar/gkac059 |
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