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The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli

The small subunit (SSU) of the ribosome of E. coli consists of a core of ribosomal RNA (rRNA) surrounded peripherally by ribosomal proteins (r-proteins). Ten of the 15 universally conserved SSU r-proteins possess nonglobular regions called extensions. The N-terminal noncanonically structured extensi...

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Autores principales: Calidas, Deepika, Lyon, Hiram, Culver, Gloria M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3923127/
https://www.ncbi.nlm.nih.gov/pubmed/24442609
http://dx.doi.org/10.1261/rna.042432.113
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author Calidas, Deepika
Lyon, Hiram
Culver, Gloria M.
author_facet Calidas, Deepika
Lyon, Hiram
Culver, Gloria M.
author_sort Calidas, Deepika
collection PubMed
description The small subunit (SSU) of the ribosome of E. coli consists of a core of ribosomal RNA (rRNA) surrounded peripherally by ribosomal proteins (r-proteins). Ten of the 15 universally conserved SSU r-proteins possess nonglobular regions called extensions. The N-terminal noncanonically structured extension of S12 traverses from the solvent to intersubunit surface of the SSU and is followed by a more C-terminal globular region that is adjacent to the decoding center of the SSU. The role of the globular region in maintaining translational fidelity is well characterized, but a role for the S12 extension in SSU structure and function is unknown. We examined the effect of stepwise truncation of the extension of S12 in SSU assembly and function in vitro and in vivo. Examination of in vitro assembly in the presence of sequential N-terminal truncated variants of S12 reveals that N-terminal deletions of greater than nine amino acids exhibit decreased tRNA-binding activity and altered 16S rRNA architecture particularly in the platform of the SSU. While wild-type S12 expressed from a plasmid can rescue a genomic deletion of the essential gene for S12, rpsl; N-terminal deletions of S12 exhibit deleterious phenotypic consequences. Partial N-terminal deletions of S12 are slow growing and cold sensitive. Strains bearing these truncations as the sole copy of S12 have increased levels of free SSUs and immature 16S rRNA as compared with the wild-type S12. These differences are hallmarks of SSU biogenesis defects, indicating that the extension of S12 plays an important role in SSU assembly.
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spelling pubmed-39231272015-03-01 The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli Calidas, Deepika Lyon, Hiram Culver, Gloria M. RNA Articles The small subunit (SSU) of the ribosome of E. coli consists of a core of ribosomal RNA (rRNA) surrounded peripherally by ribosomal proteins (r-proteins). Ten of the 15 universally conserved SSU r-proteins possess nonglobular regions called extensions. The N-terminal noncanonically structured extension of S12 traverses from the solvent to intersubunit surface of the SSU and is followed by a more C-terminal globular region that is adjacent to the decoding center of the SSU. The role of the globular region in maintaining translational fidelity is well characterized, but a role for the S12 extension in SSU structure and function is unknown. We examined the effect of stepwise truncation of the extension of S12 in SSU assembly and function in vitro and in vivo. Examination of in vitro assembly in the presence of sequential N-terminal truncated variants of S12 reveals that N-terminal deletions of greater than nine amino acids exhibit decreased tRNA-binding activity and altered 16S rRNA architecture particularly in the platform of the SSU. While wild-type S12 expressed from a plasmid can rescue a genomic deletion of the essential gene for S12, rpsl; N-terminal deletions of S12 exhibit deleterious phenotypic consequences. Partial N-terminal deletions of S12 are slow growing and cold sensitive. Strains bearing these truncations as the sole copy of S12 have increased levels of free SSUs and immature 16S rRNA as compared with the wild-type S12. These differences are hallmarks of SSU biogenesis defects, indicating that the extension of S12 plays an important role in SSU assembly. Cold Spring Harbor Laboratory Press 2014-03 /pmc/articles/PMC3923127/ /pubmed/24442609 http://dx.doi.org/10.1261/rna.042432.113 Text en © 2014 Calidas et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/3.0/ This article is distributed exclusively by the RNA Society for the first 12 months after the full-issue publication date (see http://rnajournal.cshlp.org/site/misc/terms.xhtml). After 12 months, it is available under a Creative Commons License (Attribution-NonCommercial 3.0 Unported), as described at http://creativecommons.org/licenses/by-nc/3.0/.
spellingShingle Articles
Calidas, Deepika
Lyon, Hiram
Culver, Gloria M.
The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli
title The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli
title_full The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli
title_fullStr The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli
title_full_unstemmed The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli
title_short The N-terminal extension of S12 influences small ribosomal subunit assembly in Escherichia coli
title_sort n-terminal extension of s12 influences small ribosomal subunit assembly in escherichia coli
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3923127/
https://www.ncbi.nlm.nih.gov/pubmed/24442609
http://dx.doi.org/10.1261/rna.042432.113
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