Cargando…

An S6:S18 complex inhibits translation of E. coli rpsF

More than half of the ribosomal protein operons in Escherichia coli are regulated by structures within the mRNA transcripts that interact with specific ribosomal proteins to inhibit further protein expression. This regulation is accomplished using a variety of mechanisms and the RNA structures respo...

Descripción completa

Detalles Bibliográficos
Autores principales: Babina, Arianne M., Soo, Mark W., Fu, Yang, Meyer, Michelle M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Cold Spring Harbor Laboratory Press 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4647458/
https://www.ncbi.nlm.nih.gov/pubmed/26447183
http://dx.doi.org/10.1261/rna.049544.115
_version_ 1782401103665561600
author Babina, Arianne M.
Soo, Mark W.
Fu, Yang
Meyer, Michelle M.
author_facet Babina, Arianne M.
Soo, Mark W.
Fu, Yang
Meyer, Michelle M.
author_sort Babina, Arianne M.
collection PubMed
description More than half of the ribosomal protein operons in Escherichia coli are regulated by structures within the mRNA transcripts that interact with specific ribosomal proteins to inhibit further protein expression. This regulation is accomplished using a variety of mechanisms and the RNA structures responsible for regulation are often not conserved across bacterial phyla. A widely conserved mRNA structure preceding the ribosomal protein operon containing rpsF and rpsR (encoding S6 and S18) was recently identified through comparative genomics. Examples of this RNA from both E. coli and Bacillus subtilis were shown to interact in vitro with an S6:S18 complex. In this work, we demonstrate that in E. coli, this RNA structure regulates gene expression in response to the S6:S18 complex. β-galactosidase activity from a lacZ reporter translationally fused to the 5′ UTR and first nine codons of E. coli rpsF is reduced fourfold by overexpression of a genomic fragment encoding both S6 and S18 but not by overexpression of either protein individually. Mutations to the mRNA structure, as well as to the RNA-binding site of S18 and the S6–S18 interaction surfaces of S6 and S18, are sufficient to derepress β-galactosidase activity, indicating that the S6:S18 complex is the biologically active effector. Measurement of transcript levels shows that although reporter levels do not change upon protein overexpression, levels of the native transcript are reduced fourfold, suggesting that the mRNA regulator prevents translation and this effect is amplified on the native transcript by other mechanisms.
format Online
Article
Text
id pubmed-4647458
institution National Center for Biotechnology Information
language English
publishDate 2015
publisher Cold Spring Harbor Laboratory Press
record_format MEDLINE/PubMed
spelling pubmed-46474582015-12-01 An S6:S18 complex inhibits translation of E. coli rpsF Babina, Arianne M. Soo, Mark W. Fu, Yang Meyer, Michelle M. RNA Report More than half of the ribosomal protein operons in Escherichia coli are regulated by structures within the mRNA transcripts that interact with specific ribosomal proteins to inhibit further protein expression. This regulation is accomplished using a variety of mechanisms and the RNA structures responsible for regulation are often not conserved across bacterial phyla. A widely conserved mRNA structure preceding the ribosomal protein operon containing rpsF and rpsR (encoding S6 and S18) was recently identified through comparative genomics. Examples of this RNA from both E. coli and Bacillus subtilis were shown to interact in vitro with an S6:S18 complex. In this work, we demonstrate that in E. coli, this RNA structure regulates gene expression in response to the S6:S18 complex. β-galactosidase activity from a lacZ reporter translationally fused to the 5′ UTR and first nine codons of E. coli rpsF is reduced fourfold by overexpression of a genomic fragment encoding both S6 and S18 but not by overexpression of either protein individually. Mutations to the mRNA structure, as well as to the RNA-binding site of S18 and the S6–S18 interaction surfaces of S6 and S18, are sufficient to derepress β-galactosidase activity, indicating that the S6:S18 complex is the biologically active effector. Measurement of transcript levels shows that although reporter levels do not change upon protein overexpression, levels of the native transcript are reduced fourfold, suggesting that the mRNA regulator prevents translation and this effect is amplified on the native transcript by other mechanisms. Cold Spring Harbor Laboratory Press 2015-12 /pmc/articles/PMC4647458/ /pubmed/26447183 http://dx.doi.org/10.1261/rna.049544.115 Text en © 2015 Babina et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by/4.0/ This article, published in RNA, is available under a Creative Commons License (Attribution 4.0 International), as described at http://creativecommons.org/licenses/by/4.0/.
spellingShingle Report
Babina, Arianne M.
Soo, Mark W.
Fu, Yang
Meyer, Michelle M.
An S6:S18 complex inhibits translation of E. coli rpsF
title An S6:S18 complex inhibits translation of E. coli rpsF
title_full An S6:S18 complex inhibits translation of E. coli rpsF
title_fullStr An S6:S18 complex inhibits translation of E. coli rpsF
title_full_unstemmed An S6:S18 complex inhibits translation of E. coli rpsF
title_short An S6:S18 complex inhibits translation of E. coli rpsF
title_sort s6:s18 complex inhibits translation of e. coli rpsf
topic Report
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4647458/
https://www.ncbi.nlm.nih.gov/pubmed/26447183
http://dx.doi.org/10.1261/rna.049544.115
work_keys_str_mv AT babinaariannem ans6s18complexinhibitstranslationofecolirpsf
AT soomarkw ans6s18complexinhibitstranslationofecolirpsf
AT fuyang ans6s18complexinhibitstranslationofecolirpsf
AT meyermichellem ans6s18complexinhibitstranslationofecolirpsf
AT babinaariannem s6s18complexinhibitstranslationofecolirpsf
AT soomarkw s6s18complexinhibitstranslationofecolirpsf
AT fuyang s6s18complexinhibitstranslationofecolirpsf
AT meyermichellem s6s18complexinhibitstranslationofecolirpsf