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Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663)
Small regulatory RNAs (sRNAs) play an important role for posttranscriptional gene regulation in bacteria. sRNAs recognize their target messenger RNAs (mRNAs) by base-pairing, which is often facilitated by interactions with the bacterial RNA-binding proteins Hfq or ProQ. The FinO/ProQ RNA-binding pro...
Autores principales: | , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Cold Spring Harbor Laboratory Press
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7668265/ https://www.ncbi.nlm.nih.gov/pubmed/32989045 http://dx.doi.org/10.1261/rna.077354.120 |
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author | Immer, Carina Hacker, Carolin Wöhnert, Jens |
author_facet | Immer, Carina Hacker, Carolin Wöhnert, Jens |
author_sort | Immer, Carina |
collection | PubMed |
description | Small regulatory RNAs (sRNAs) play an important role for posttranscriptional gene regulation in bacteria. sRNAs recognize their target messenger RNAs (mRNAs) by base-pairing, which is often facilitated by interactions with the bacterial RNA-binding proteins Hfq or ProQ. The FinO/ProQ RNA-binding protein domain was first discovered in the bacterial repressor of conjugation, FinO. Since then, the functional role of FinO/ProQ-like proteins in posttranscriptional gene regulation was extensively studied in particular in the enterobacteria E. coli and Salmonella enterica and a wide range of sRNA-targets was identified for these proteins. In addition, enterobacterial ProQ homologs also recognize and protect the 3′-ends of a number of mRNAs from exonucleolytic degradation. However, the RNA-binding properties of FinO/ProQ proteins with regard to the recognition of different RNA targets are not yet fully understood. Here, we present the solution NMR structure of the so far functionally uncharacterized ProQ homolog Lpp1663 from Legionella pneumophila as a newly confirmed member and a minimal model system of the FinO/ProQ protein family. In addition, we characterize the RNA-binding preferences of Lpp1663 with high resolution NMR spectroscopy and isothermal titration calorimetry (ITC). Our results suggest a binding preference for single-stranded uridine-rich RNAs in the vicinity of stable stem–loop structures. According to chemical shift perturbation experiments, the single-stranded U-rich RNAs interact mainly with a conserved RNA-binding surface on the concave site of Lpp1663. |
format | Online Article Text |
id | pubmed-7668265 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Cold Spring Harbor Laboratory Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-76682652021-12-01 Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663) Immer, Carina Hacker, Carolin Wöhnert, Jens RNA Article Small regulatory RNAs (sRNAs) play an important role for posttranscriptional gene regulation in bacteria. sRNAs recognize their target messenger RNAs (mRNAs) by base-pairing, which is often facilitated by interactions with the bacterial RNA-binding proteins Hfq or ProQ. The FinO/ProQ RNA-binding protein domain was first discovered in the bacterial repressor of conjugation, FinO. Since then, the functional role of FinO/ProQ-like proteins in posttranscriptional gene regulation was extensively studied in particular in the enterobacteria E. coli and Salmonella enterica and a wide range of sRNA-targets was identified for these proteins. In addition, enterobacterial ProQ homologs also recognize and protect the 3′-ends of a number of mRNAs from exonucleolytic degradation. However, the RNA-binding properties of FinO/ProQ proteins with regard to the recognition of different RNA targets are not yet fully understood. Here, we present the solution NMR structure of the so far functionally uncharacterized ProQ homolog Lpp1663 from Legionella pneumophila as a newly confirmed member and a minimal model system of the FinO/ProQ protein family. In addition, we characterize the RNA-binding preferences of Lpp1663 with high resolution NMR spectroscopy and isothermal titration calorimetry (ITC). Our results suggest a binding preference for single-stranded uridine-rich RNAs in the vicinity of stable stem–loop structures. According to chemical shift perturbation experiments, the single-stranded U-rich RNAs interact mainly with a conserved RNA-binding surface on the concave site of Lpp1663. Cold Spring Harbor Laboratory Press 2020-12 /pmc/articles/PMC7668265/ /pubmed/32989045 http://dx.doi.org/10.1261/rna.077354.120 Text en © 2020 Immer et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society http://creativecommons.org/licenses/by-nc/4.0/ 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 4.0 International), as described at http://creativecommons.org/licenses/by-nc/4.0/. |
spellingShingle | Article Immer, Carina Hacker, Carolin Wöhnert, Jens Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663) |
title | Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663) |
title_full | Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663) |
title_fullStr | Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663) |
title_full_unstemmed | Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663) |
title_short | Solution structure and RNA-binding of a minimal ProQ-homolog from Legionella pneumophila (Lpp1663) |
title_sort | solution structure and rna-binding of a minimal proq-homolog from legionella pneumophila (lpp1663) |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7668265/ https://www.ncbi.nlm.nih.gov/pubmed/32989045 http://dx.doi.org/10.1261/rna.077354.120 |
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