Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>

ABSTRACT Elongation factor P (EF-P) is a universally conserved bacterial translation factor homologous to eukaryotic/archaeal initiation factor 5A. In Salmonella, deletion of the efp gene results in pleiotropic phenotypes, including increased susceptibility to numerous cellular stressors. Only a lim...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Steven J. Hersch, Mengchi Wang, S. Betty Zou, Kyung-Mee Moon, Leonard J. Foster, Michael Ibba, William Wiley Navarre
Formato:	article
Lenguaje:	EN
Publicado:	American Society for Microbiology 2013
Materias:	Microbiology QR1-502
Acceso en línea:	https://doaj.org/article/9797cfa4e47a47b998470c2ed5304fda
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:9797cfa4e47a47b998470c2ed5304fda
record_format	dspace
spelling	oai:doaj.org-article:9797cfa4e47a47b998470c2ed5304fda2021-11-15T15:40:28ZDivergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>10.1128/mBio.00180-132150-7511https://doaj.org/article/9797cfa4e47a47b998470c2ed5304fda2013-05-01T00:00:00Zhttps://journals.asm.org/doi/10.1128/mBio.00180-13https://doaj.org/toc/2150-7511ABSTRACT Elongation factor P (EF-P) is a universally conserved bacterial translation factor homologous to eukaryotic/archaeal initiation factor 5A. In Salmonella, deletion of the efp gene results in pleiotropic phenotypes, including increased susceptibility to numerous cellular stressors. Only a limited number of proteins are affected by the loss of EF-P, and it has recently been determined that EF-P plays a critical role in rescuing ribosomes stalled at PPP and PPG peptide sequences. Here we present an unbiased in vivo investigation of the specific targets of EF-P by employing stable isotope labeling of amino acids in cell culture (SILAC) to compare the proteomes of wild-type and efp mutant Salmonella. We found that metabolic and motility genes are prominent among the subset of proteins with decreased production in the Δefp mutant. Furthermore, particular tripeptide motifs are statistically overrepresented among the proteins downregulated in efp mutant strains. These include both PPP and PPG but also additional motifs, such as APP and YIRYIR, which were confirmed to induce EF-P dependence by a translational fusion assay. Notably, we found that many proteins containing polyproline motifs are not misregulated in an EF-P-deficient background, suggesting that the factors that govern EF-P-mediated regulation are complex. Finally, we analyzed the specific region of the PoxB protein that is modulated by EF-P and found that mutation of any residue within a specific GSCGPG sequence eliminates the requirement for EF-P. This work expands the known repertoire of EF-P target motifs and implicates factors beyond polyproline motifs that are required for EF-P-mediated regulation. IMPORTANCE Bacterial cells regulate gene expression at several points during and after transcription. During protein synthesis, for example, factors can interact with the ribosome to influence the production of specific proteins. Bacterial elongation factor P (EF-P) is a protein that facilitates the synthesis of proteins that contain polyproline motifs by preventing the ribosome from stalling. Bacterial cells that lack EF-P are viable but are sensitive to a large number of stress conditions. In this study, a global analysis of protein synthesis revealed that EF-P regulates many more proteins in the cell than predicted based solely on the prevalence of polyproline motifs. Several new EF-P-regulated motifs were uncovered, thereby providing a more complete picture of how this critical factor influences the cell’s response to stress at the level of protein synthesis.Steven J. HerschMengchi WangS. Betty ZouKyung-Mee MoonLeonard J. FosterMichael IbbaWilliam Wiley NavarreAmerican Society for MicrobiologyarticleMicrobiologyQR1-502ENmBio, Vol 4, Iss 2 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Microbiology QR1-502
spellingShingle	Microbiology QR1-502 Steven J. Hersch Mengchi Wang S. Betty Zou Kyung-Mee Moon Leonard J. Foster Michael Ibba William Wiley Navarre Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>
description	ABSTRACT Elongation factor P (EF-P) is a universally conserved bacterial translation factor homologous to eukaryotic/archaeal initiation factor 5A. In Salmonella, deletion of the efp gene results in pleiotropic phenotypes, including increased susceptibility to numerous cellular stressors. Only a limited number of proteins are affected by the loss of EF-P, and it has recently been determined that EF-P plays a critical role in rescuing ribosomes stalled at PPP and PPG peptide sequences. Here we present an unbiased in vivo investigation of the specific targets of EF-P by employing stable isotope labeling of amino acids in cell culture (SILAC) to compare the proteomes of wild-type and efp mutant Salmonella. We found that metabolic and motility genes are prominent among the subset of proteins with decreased production in the Δefp mutant. Furthermore, particular tripeptide motifs are statistically overrepresented among the proteins downregulated in efp mutant strains. These include both PPP and PPG but also additional motifs, such as APP and YIRYIR, which were confirmed to induce EF-P dependence by a translational fusion assay. Notably, we found that many proteins containing polyproline motifs are not misregulated in an EF-P-deficient background, suggesting that the factors that govern EF-P-mediated regulation are complex. Finally, we analyzed the specific region of the PoxB protein that is modulated by EF-P and found that mutation of any residue within a specific GSCGPG sequence eliminates the requirement for EF-P. This work expands the known repertoire of EF-P target motifs and implicates factors beyond polyproline motifs that are required for EF-P-mediated regulation. IMPORTANCE Bacterial cells regulate gene expression at several points during and after transcription. During protein synthesis, for example, factors can interact with the ribosome to influence the production of specific proteins. Bacterial elongation factor P (EF-P) is a protein that facilitates the synthesis of proteins that contain polyproline motifs by preventing the ribosome from stalling. Bacterial cells that lack EF-P are viable but are sensitive to a large number of stress conditions. In this study, a global analysis of protein synthesis revealed that EF-P regulates many more proteins in the cell than predicted based solely on the prevalence of polyproline motifs. Several new EF-P-regulated motifs were uncovered, thereby providing a more complete picture of how this critical factor influences the cell’s response to stress at the level of protein synthesis.
format	article
author	Steven J. Hersch Mengchi Wang S. Betty Zou Kyung-Mee Moon Leonard J. Foster Michael Ibba William Wiley Navarre
author_facet	Steven J. Hersch Mengchi Wang S. Betty Zou Kyung-Mee Moon Leonard J. Foster Michael Ibba William Wiley Navarre
author_sort	Steven J. Hersch
title	Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>
title_short	Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>
title_full	Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>
title_fullStr	Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>
title_full_unstemmed	Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>
title_sort	divergent protein motifs direct elongation factor p-mediated translational regulation in <named-content content-type="genus-species">salmonella enterica</named-content> and <named-content content-type="genus-species">escherichia coli</named-content>
publisher	American Society for Microbiology
publishDate	2013
url	https://doaj.org/article/9797cfa4e47a47b998470c2ed5304fda
work_keys_str_mv	AT stevenjhersch divergentproteinmotifsdirectelongationfactorpmediatedtranslationalregulationinnamedcontentcontenttypegenusspeciessalmonellaentericanamedcontentandnamedcontentcontenttypegenusspeciesescherichiacolinamedcontent AT mengchiwang divergentproteinmotifsdirectelongationfactorpmediatedtranslationalregulationinnamedcontentcontenttypegenusspeciessalmonellaentericanamedcontentandnamedcontentcontenttypegenusspeciesescherichiacolinamedcontent AT sbettyzou divergentproteinmotifsdirectelongationfactorpmediatedtranslationalregulationinnamedcontentcontenttypegenusspeciessalmonellaentericanamedcontentandnamedcontentcontenttypegenusspeciesescherichiacolinamedcontent AT kyungmeemoon divergentproteinmotifsdirectelongationfactorpmediatedtranslationalregulationinnamedcontentcontenttypegenusspeciessalmonellaentericanamedcontentandnamedcontentcontenttypegenusspeciesescherichiacolinamedcontent AT leonardjfoster divergentproteinmotifsdirectelongationfactorpmediatedtranslationalregulationinnamedcontentcontenttypegenusspeciessalmonellaentericanamedcontentandnamedcontentcontenttypegenusspeciesescherichiacolinamedcontent AT michaelibba divergentproteinmotifsdirectelongationfactorpmediatedtranslationalregulationinnamedcontentcontenttypegenusspeciessalmonellaentericanamedcontentandnamedcontentcontenttypegenusspeciesescherichiacolinamedcontent AT williamwileynavarre divergentproteinmotifsdirectelongationfactorpmediatedtranslationalregulationinnamedcontentcontenttypegenusspeciessalmonellaentericanamedcontentandnamedcontentcontenttypegenusspeciesescherichiacolinamedcontent
_version_	1718427717734498304

Divergent Protein Motifs Direct Elongation Factor P-Mediated Translational Regulation in <named-content content-type="genus-species">Salmonella enterica</named-content> and <named-content content-type="genus-species">Escherichia coli</named-content>

Ejemplares similares