Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>

ABSTRACT The eukaryotic ribosome consists of a small (40S) and a large (60S) subunit. Rps26 is one of the essential ribosomal proteins of the 40S subunit and is encoded by two almost identical genes, RPS26a and RPS26b. Previous studies demonstrated that Rps26 interacts with the 5′ untranslated regio...

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Autores principales:	Alexander Belyy, Nadezhda Levanova, Irina Tabakova, Sabine Rospert, Yury Belyi
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2016
Materias:	40S subunit Saccharomyces cerevisiae eukaryote-specific motif mutagenesis ribosomal protein ribosome assembly Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/c0261629fa684336a4bdcf5ad3286c92
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spelling	oai:doaj.org-article:c0261629fa684336a4bdcf5ad3286c922021-11-15T15:21:38ZRibosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>10.1128/mSphere.00109-152379-5042https://doaj.org/article/c0261629fa684336a4bdcf5ad3286c922016-02-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mSphere.00109-15https://doaj.org/toc/2379-5042ABSTRACT The eukaryotic ribosome consists of a small (40S) and a large (60S) subunit. Rps26 is one of the essential ribosomal proteins of the 40S subunit and is encoded by two almost identical genes, RPS26a and RPS26b. Previous studies demonstrated that Rps26 interacts with the 5′ untranslated region of mRNA via the eukaryote-specific 62-YXXPKXYXK-70 (Y62–K70) motif. Those observations suggested that this peptide within Rps26 might play an important and specific role during translation initiation. By using alanine-scanning mutagenesis and engineered strains of the yeast Saccharomyces cerevisiae, we found that single amino acid substitutions within the Y62–K70 motif of Rps26 did not affect the in vivo function of the protein. In contrast, complete deletion of the Y62–K70 segment was lethal. The simultaneous replacement of five conserved residues within the Y62–K70 segment by alanines resulted in growth defects under stress conditions and produced distinct changes in polysome profiles that were indicative of the accumulation of free 60S subunits. Human Rps26 (Rps26-Hs), which displays significant homology with yeast Rps26, supported the growth of an S. cerevisiae Δrps26a Δrps26b strain. However, the Δrps26a Δrps26b double deletion strain expressing Rps26-Hs displayed substantial growth defects and an altered ratio of 40S/60S ribosomal subunits. The combined data strongly suggest that the eukaryote-specific motif within Rps26 does not play a specific role in translation initiation. Rather, the data indicate that Rps26 as a whole is necessary for proper assembly of the 40S subunit and the 80S ribosome in yeast. IMPORTANCE Rps26 is an essential protein of the eukaryotic small ribosomal subunit. Previous experiments demonstrated an interaction between the eukaryote-specific Y62–K70 segment of Rps26 and the 5′ untranslated region of mRNA. The data suggested a specific role of the Y62–K70 motif during translation initiation. Here, we report that single-site substitutions within the Y62–K70 peptide did not affect the growth of engineered yeast strains, arguing against its having a critical role during translation initiation via specific interactions with the 5′ untranslated region of mRNA molecules. Only the simultaneous replacement of five conserved residues within the Y62–K70 fragment or the replacement of the yeast protein with the human homolog resulted in growth defects and caused significant changes in polysome profiles. The results expand our knowledge of ribosomal protein function and suggest a role of Rps26 during ribosome assembly in yeast.Alexander BelyyNadezhda LevanovaIrina TabakovaSabine RospertYury BelyiAmerican Society for Microbiologyarticle40S subunitSaccharomyces cerevisiaeeukaryote-specific motifmutagenesisribosomal proteinribosome assemblyMicrobiologyQR1-502ENmSphere, Vol 1, Iss 1 (2016)
institution	DOAJ
collection	DOAJ
language	EN
topic	40S subunit Saccharomyces cerevisiae eukaryote-specific motif mutagenesis ribosomal protein ribosome assembly Microbiology QR1-502
spellingShingle	40S subunit Saccharomyces cerevisiae eukaryote-specific motif mutagenesis ribosomal protein ribosome assembly Microbiology QR1-502 Alexander Belyy Nadezhda Levanova Irina Tabakova Sabine Rospert Yury Belyi Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>
description	ABSTRACT The eukaryotic ribosome consists of a small (40S) and a large (60S) subunit. Rps26 is one of the essential ribosomal proteins of the 40S subunit and is encoded by two almost identical genes, RPS26a and RPS26b. Previous studies demonstrated that Rps26 interacts with the 5′ untranslated region of mRNA via the eukaryote-specific 62-YXXPKXYXK-70 (Y62–K70) motif. Those observations suggested that this peptide within Rps26 might play an important and specific role during translation initiation. By using alanine-scanning mutagenesis and engineered strains of the yeast Saccharomyces cerevisiae, we found that single amino acid substitutions within the Y62–K70 motif of Rps26 did not affect the in vivo function of the protein. In contrast, complete deletion of the Y62–K70 segment was lethal. The simultaneous replacement of five conserved residues within the Y62–K70 segment by alanines resulted in growth defects under stress conditions and produced distinct changes in polysome profiles that were indicative of the accumulation of free 60S subunits. Human Rps26 (Rps26-Hs), which displays significant homology with yeast Rps26, supported the growth of an S. cerevisiae Δrps26a Δrps26b strain. However, the Δrps26a Δrps26b double deletion strain expressing Rps26-Hs displayed substantial growth defects and an altered ratio of 40S/60S ribosomal subunits. The combined data strongly suggest that the eukaryote-specific motif within Rps26 does not play a specific role in translation initiation. Rather, the data indicate that Rps26 as a whole is necessary for proper assembly of the 40S subunit and the 80S ribosome in yeast. IMPORTANCE Rps26 is an essential protein of the eukaryotic small ribosomal subunit. Previous experiments demonstrated an interaction between the eukaryote-specific Y62–K70 segment of Rps26 and the 5′ untranslated region of mRNA. The data suggested a specific role of the Y62–K70 motif during translation initiation. Here, we report that single-site substitutions within the Y62–K70 peptide did not affect the growth of engineered yeast strains, arguing against its having a critical role during translation initiation via specific interactions with the 5′ untranslated region of mRNA molecules. Only the simultaneous replacement of five conserved residues within the Y62–K70 fragment or the replacement of the yeast protein with the human homolog resulted in growth defects and caused significant changes in polysome profiles. The results expand our knowledge of ribosomal protein function and suggest a role of Rps26 during ribosome assembly in yeast.
format	article
author	Alexander Belyy Nadezhda Levanova Irina Tabakova Sabine Rospert Yury Belyi
author_facet	Alexander Belyy Nadezhda Levanova Irina Tabakova Sabine Rospert Yury Belyi
author_sort	Alexander Belyy
title	Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>
title_short	Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>
title_full	Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>
title_fullStr	Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>
title_full_unstemmed	Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>
title_sort	ribosomal protein rps26 influences 80s ribosome assembly in <named-content content-type="genus-species">saccharomyces cerevisiae</named-content>
publisher	American Society for Microbiology
publishDate	2016
url	https://doaj.org/article/c0261629fa684336a4bdcf5ad3286c92
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Ribosomal Protein Rps26 Influences 80S Ribosome Assembly in <named-content content-type="genus-species">Saccharomyces cerevisiae</named-content>

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