Cargando…

Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation

Using sample-matched transcriptomics and proteomics measurements it is now possible to begin to understand the impact of post-transcriptional regulatory programs in Enterobacteria. In bacteria post-transcriptional regulation is mediated by relatively few identified RNA-binding protein factors includ...

Descripción completa

Detalles Bibliográficos
Autores principales: Ansong, Charles, Yoon, Hyunjin, Porwollik, Steffen, Mottaz-Brewer, Heather, Petritis, Brianne O., Jaitly, Navdeep, Adkins, Joshua N., McClelland, Michael, Heffron, Fred, Smith, Richard D.
Formato: Texto
Lenguaje:English
Publicado: Public Library of Science 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2652828/
https://www.ncbi.nlm.nih.gov/pubmed/19277208
http://dx.doi.org/10.1371/journal.pone.0004809
_version_ 1782165254567886848
author Ansong, Charles
Yoon, Hyunjin
Porwollik, Steffen
Mottaz-Brewer, Heather
Petritis, Brianne O.
Jaitly, Navdeep
Adkins, Joshua N.
McClelland, Michael
Heffron, Fred
Smith, Richard D.
author_facet Ansong, Charles
Yoon, Hyunjin
Porwollik, Steffen
Mottaz-Brewer, Heather
Petritis, Brianne O.
Jaitly, Navdeep
Adkins, Joshua N.
McClelland, Michael
Heffron, Fred
Smith, Richard D.
author_sort Ansong, Charles
collection PubMed
description Using sample-matched transcriptomics and proteomics measurements it is now possible to begin to understand the impact of post-transcriptional regulatory programs in Enterobacteria. In bacteria post-transcriptional regulation is mediated by relatively few identified RNA-binding protein factors including CsrA, Hfq and SmpB. A mutation in any one of these three genes, csrA, hfq, and smpB, in Salmonella is attenuated for mouse virulence and unable to survive in macrophages. CsrA has a clearly defined specificity based on binding to a specific mRNA sequence to inhibit translation. However, the proteins regulated by Hfq and SmpB are not as clearly defined. Previous work identified proteins regulated by hfq using purification of the RNA-protein complex with direct sequencing of the bound RNAs and found binding to a surprisingly large number of transcripts. In this report we have used global proteomics to directly identify proteins regulated by Hfq or SmpB by comparing protein abundance in the parent and isogenic hfq or smpB mutant. From these same samples we also prepared RNA for microarray analysis to determine if alteration of protein expression was mediated post-transcriptionally. Samples were analyzed from bacteria grown under four different conditions; two laboratory conditions and two that are thought to mimic the intracellular environment. We show that mutants of hfq and smpB directly or indirectly modulate at least 20% and 4% of all possible Salmonella proteins, respectively, with limited correlation between transcription and protein expression. These proteins represent a broad spectrum of Salmonella proteins required for many biological processes including host cell invasion, motility, central metabolism, LPS biosynthesis, two-component regulatory systems, and fatty acid metabolism. Our results represent one of the first global analyses of post-transcriptional regulons in any organism and suggest that regulation at the translational level is widespread and plays an important role in virulence regulation and environmental adaptation for Salmonella.
format Text
id pubmed-2652828
institution National Center for Biotechnology Information
language English
publishDate 2009
publisher Public Library of Science
record_format MEDLINE/PubMed
spelling pubmed-26528282009-03-11 Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation Ansong, Charles Yoon, Hyunjin Porwollik, Steffen Mottaz-Brewer, Heather Petritis, Brianne O. Jaitly, Navdeep Adkins, Joshua N. McClelland, Michael Heffron, Fred Smith, Richard D. PLoS One Research Article Using sample-matched transcriptomics and proteomics measurements it is now possible to begin to understand the impact of post-transcriptional regulatory programs in Enterobacteria. In bacteria post-transcriptional regulation is mediated by relatively few identified RNA-binding protein factors including CsrA, Hfq and SmpB. A mutation in any one of these three genes, csrA, hfq, and smpB, in Salmonella is attenuated for mouse virulence and unable to survive in macrophages. CsrA has a clearly defined specificity based on binding to a specific mRNA sequence to inhibit translation. However, the proteins regulated by Hfq and SmpB are not as clearly defined. Previous work identified proteins regulated by hfq using purification of the RNA-protein complex with direct sequencing of the bound RNAs and found binding to a surprisingly large number of transcripts. In this report we have used global proteomics to directly identify proteins regulated by Hfq or SmpB by comparing protein abundance in the parent and isogenic hfq or smpB mutant. From these same samples we also prepared RNA for microarray analysis to determine if alteration of protein expression was mediated post-transcriptionally. Samples were analyzed from bacteria grown under four different conditions; two laboratory conditions and two that are thought to mimic the intracellular environment. We show that mutants of hfq and smpB directly or indirectly modulate at least 20% and 4% of all possible Salmonella proteins, respectively, with limited correlation between transcription and protein expression. These proteins represent a broad spectrum of Salmonella proteins required for many biological processes including host cell invasion, motility, central metabolism, LPS biosynthesis, two-component regulatory systems, and fatty acid metabolism. Our results represent one of the first global analyses of post-transcriptional regulons in any organism and suggest that regulation at the translational level is widespread and plays an important role in virulence regulation and environmental adaptation for Salmonella. Public Library of Science 2009-03-11 /pmc/articles/PMC2652828/ /pubmed/19277208 http://dx.doi.org/10.1371/journal.pone.0004809 Text en This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose. https://creativecommons.org/publicdomain/zero/1.0/ This is an open-access article distributed under the terms of the Creative Commons Public Domain declaration, which stipulates that, once placed in the public domain, this work may be freely reproduced, distributed, transmitted, modified, built upon, or otherwise used by anyone for any lawful purpose.
spellingShingle Research Article
Ansong, Charles
Yoon, Hyunjin
Porwollik, Steffen
Mottaz-Brewer, Heather
Petritis, Brianne O.
Jaitly, Navdeep
Adkins, Joshua N.
McClelland, Michael
Heffron, Fred
Smith, Richard D.
Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation
title Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation
title_full Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation
title_fullStr Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation
title_full_unstemmed Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation
title_short Global Systems-Level Analysis of Hfq and SmpB Deletion Mutants in Salmonella: Implications for Virulence and Global Protein Translation
title_sort global systems-level analysis of hfq and smpb deletion mutants in salmonella: implications for virulence and global protein translation
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2652828/
https://www.ncbi.nlm.nih.gov/pubmed/19277208
http://dx.doi.org/10.1371/journal.pone.0004809
work_keys_str_mv AT ansongcharles globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT yoonhyunjin globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT porwolliksteffen globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT mottazbrewerheather globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT petritisbrianneo globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT jaitlynavdeep globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT adkinsjoshuan globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT mcclellandmichael globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT heffronfred globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation
AT smithrichardd globalsystemslevelanalysisofhfqandsmpbdeletionmutantsinsalmonellaimplicationsforvirulenceandglobalproteintranslation