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The Hfq regulon of Neisseria meningitidis

The conserved RNA‐binding protein, Hfq, has multiple regulatory roles within the prokaryotic cell, including promoting stable duplex formation between small RNAs and mRNAs, and thus hfq deletion mutants have pleiotropic phenotypes. Previous proteome and transcriptome studies of Neisseria meningitidi...

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Autores principales: Huis in ‘t Veld, Robert A. G., Kramer, Gertjan, van der Ende, Arie, Speijer, Dave, Pannekoek, Yvonne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5458458/
https://www.ncbi.nlm.nih.gov/pubmed/28593133
http://dx.doi.org/10.1002/2211-5463.12218
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author Huis in ‘t Veld, Robert A. G.
Kramer, Gertjan
van der Ende, Arie
Speijer, Dave
Pannekoek, Yvonne
author_facet Huis in ‘t Veld, Robert A. G.
Kramer, Gertjan
van der Ende, Arie
Speijer, Dave
Pannekoek, Yvonne
author_sort Huis in ‘t Veld, Robert A. G.
collection PubMed
description The conserved RNA‐binding protein, Hfq, has multiple regulatory roles within the prokaryotic cell, including promoting stable duplex formation between small RNAs and mRNAs, and thus hfq deletion mutants have pleiotropic phenotypes. Previous proteome and transcriptome studies of Neisseria meningitidis have generated limited insight into differential gene expression due to Hfq loss. In this study, reversed‐phase liquid chromatography combined with data‐independent alternate scanning mass spectrometry (LC‐MS(E)) was utilized for rapid high‐resolution quantitative proteomic analysis to further elucidate the differentially expressed proteome of a meningococcal hfq deletion mutant. Whole‐cell lysates of N. meningitidis serogroup B H44/76 wild‐type (wt) and H44/76Δhfq (Δhfq) grown in liquid growth medium were subjected to tryptic digestion. The resulting peptide mixtures were separated by liquid chromatography (LC) prior to analysis by mass spectrometry (MS(E)). Differential expression was analyzed by Student's t‐test with control for false discovery rate (FDR). Reliable quantitation of relative expression comparing wt and Δhfq was achieved with 506 proteins (20%). Upon FDR control at q ≤ 0.05, 48 up‐ and 59 downregulated proteins were identified. From these, 81 were identified as novel Hfq‐regulated candidates, while 15 proteins were previously found by SDS/PAGE/MS and 24 with microarray analyses. Thus, using LC‐MS(E) we have expanded the repertoire of Hfq‐regulated proteins. In conjunction with previous studies, a comprehensive network of Hfq‐regulated proteins was constructed and differentially expressed proteins were found to be involved in a large variety of cellular processes. The results and comparisons with other gram‐negative model systems, suggest still unidentified sRNA analogs in N. meningitidis.
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spelling pubmed-54584582017-06-07 The Hfq regulon of Neisseria meningitidis Huis in ‘t Veld, Robert A. G. Kramer, Gertjan van der Ende, Arie Speijer, Dave Pannekoek, Yvonne FEBS Open Bio Research Articles The conserved RNA‐binding protein, Hfq, has multiple regulatory roles within the prokaryotic cell, including promoting stable duplex formation between small RNAs and mRNAs, and thus hfq deletion mutants have pleiotropic phenotypes. Previous proteome and transcriptome studies of Neisseria meningitidis have generated limited insight into differential gene expression due to Hfq loss. In this study, reversed‐phase liquid chromatography combined with data‐independent alternate scanning mass spectrometry (LC‐MS(E)) was utilized for rapid high‐resolution quantitative proteomic analysis to further elucidate the differentially expressed proteome of a meningococcal hfq deletion mutant. Whole‐cell lysates of N. meningitidis serogroup B H44/76 wild‐type (wt) and H44/76Δhfq (Δhfq) grown in liquid growth medium were subjected to tryptic digestion. The resulting peptide mixtures were separated by liquid chromatography (LC) prior to analysis by mass spectrometry (MS(E)). Differential expression was analyzed by Student's t‐test with control for false discovery rate (FDR). Reliable quantitation of relative expression comparing wt and Δhfq was achieved with 506 proteins (20%). Upon FDR control at q ≤ 0.05, 48 up‐ and 59 downregulated proteins were identified. From these, 81 were identified as novel Hfq‐regulated candidates, while 15 proteins were previously found by SDS/PAGE/MS and 24 with microarray analyses. Thus, using LC‐MS(E) we have expanded the repertoire of Hfq‐regulated proteins. In conjunction with previous studies, a comprehensive network of Hfq‐regulated proteins was constructed and differentially expressed proteins were found to be involved in a large variety of cellular processes. The results and comparisons with other gram‐negative model systems, suggest still unidentified sRNA analogs in N. meningitidis. John Wiley and Sons Inc. 2017-04-25 /pmc/articles/PMC5458458/ /pubmed/28593133 http://dx.doi.org/10.1002/2211-5463.12218 Text en © 2017 The Authors. Published by FEBS Press and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution (http://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Huis in ‘t Veld, Robert A. G.
Kramer, Gertjan
van der Ende, Arie
Speijer, Dave
Pannekoek, Yvonne
The Hfq regulon of Neisseria meningitidis
title The Hfq regulon of Neisseria meningitidis
title_full The Hfq regulon of Neisseria meningitidis
title_fullStr The Hfq regulon of Neisseria meningitidis
title_full_unstemmed The Hfq regulon of Neisseria meningitidis
title_short The Hfq regulon of Neisseria meningitidis
title_sort hfq regulon of neisseria meningitidis
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5458458/
https://www.ncbi.nlm.nih.gov/pubmed/28593133
http://dx.doi.org/10.1002/2211-5463.12218
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