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Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ

The FinO-domain-protein ProQ is an RNA-binding protein that has been known to play a role in osmoregulation in proteobacteria. Recently, ProQ has been shown to act as a global RNA-binding protein in Salmonella and Escherichia coli, binding to dozens of small RNAs (sRNAs) and messenger RNAs (mRNAs) t...

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Autores principales: Pandey, Smriti, Gravel, Chandra M, Stockert, Oliver M, Wang, Clara D, Hegner, Courtney L, LeBlanc, Hannah, Berry, Katherine E
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Oxford University Press 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192607/
https://www.ncbi.nlm.nih.gov/pubmed/32170306
http://dx.doi.org/10.1093/nar/gkaa144
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author Pandey, Smriti
Gravel, Chandra M
Stockert, Oliver M
Wang, Clara D
Hegner, Courtney L
LeBlanc, Hannah
Berry, Katherine E
author_facet Pandey, Smriti
Gravel, Chandra M
Stockert, Oliver M
Wang, Clara D
Hegner, Courtney L
LeBlanc, Hannah
Berry, Katherine E
author_sort Pandey, Smriti
collection PubMed
description The FinO-domain-protein ProQ is an RNA-binding protein that has been known to play a role in osmoregulation in proteobacteria. Recently, ProQ has been shown to act as a global RNA-binding protein in Salmonella and Escherichia coli, binding to dozens of small RNAs (sRNAs) and messenger RNAs (mRNAs) to regulate mRNA-expression levels through interactions with both 5′ and 3′ untranslated regions (UTRs). Despite excitement around ProQ as a novel global RNA-binding protein, and its potential to serve as a matchmaking RNA chaperone, significant gaps remain in our understanding of the molecular mechanisms ProQ uses to interact with RNA. In order to apply the tools of molecular genetics to this question, we have adapted a bacterial three-hybrid (B3H) assay to detect ProQ’s interactions with target RNAs. Using domain truncations, site-directed mutagenesis and an unbiased forward genetic screen, we have identified a group of highly conserved residues on ProQ’s NTD as the primary face for in vivo recognition of two RNAs, and propose that the NTD structure serves as an electrostatic scaffold to recognize the shape of an RNA duplex.
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spelling pubmed-71926072020-05-06 Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ Pandey, Smriti Gravel, Chandra M Stockert, Oliver M Wang, Clara D Hegner, Courtney L LeBlanc, Hannah Berry, Katherine E Nucleic Acids Res RNA and RNA-protein complexes The FinO-domain-protein ProQ is an RNA-binding protein that has been known to play a role in osmoregulation in proteobacteria. Recently, ProQ has been shown to act as a global RNA-binding protein in Salmonella and Escherichia coli, binding to dozens of small RNAs (sRNAs) and messenger RNAs (mRNAs) to regulate mRNA-expression levels through interactions with both 5′ and 3′ untranslated regions (UTRs). Despite excitement around ProQ as a novel global RNA-binding protein, and its potential to serve as a matchmaking RNA chaperone, significant gaps remain in our understanding of the molecular mechanisms ProQ uses to interact with RNA. In order to apply the tools of molecular genetics to this question, we have adapted a bacterial three-hybrid (B3H) assay to detect ProQ’s interactions with target RNAs. Using domain truncations, site-directed mutagenesis and an unbiased forward genetic screen, we have identified a group of highly conserved residues on ProQ’s NTD as the primary face for in vivo recognition of two RNAs, and propose that the NTD structure serves as an electrostatic scaffold to recognize the shape of an RNA duplex. Oxford University Press 2020-05-07 2020-03-14 /pmc/articles/PMC7192607/ /pubmed/32170306 http://dx.doi.org/10.1093/nar/gkaa144 Text en © The Author(s) 2020. Published by Oxford University Press on behalf of Nucleic Acids Research. http://creativecommons.org/licenses/by/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle RNA and RNA-protein complexes
Pandey, Smriti
Gravel, Chandra M
Stockert, Oliver M
Wang, Clara D
Hegner, Courtney L
LeBlanc, Hannah
Berry, Katherine E
Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ
title Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ
title_full Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ
title_fullStr Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ
title_full_unstemmed Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ
title_short Genetic identification of the functional surface for RNA binding by Escherichia coli ProQ
title_sort genetic identification of the functional surface for rna binding by escherichia coli proq
topic RNA and RNA-protein complexes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7192607/
https://www.ncbi.nlm.nih.gov/pubmed/32170306
http://dx.doi.org/10.1093/nar/gkaa144
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