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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...

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Autores principales: Seffouh, Amal, Trahan, Chirstian, Wasi, Tanzila, Jain, Nikhil, Basu, Kaustuv, Britton, Robert A, Oeffinger, Marlene, Ortega, Joaquin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2022
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.
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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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