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A census of RNA/protein complexes in a model Gram-positive bacterium reveals exonuclease-mediated sRNA activation in competence regulation

ABSTRACT: 22 total proteome samples from S. pneumoniae wild type resolved on a glycerol gradient resulting in 20 fractions and a pellet fraction. A lysate sample is given as input control.

INSTRUMENT(S): LTQ Orbitrap Velos, Orbitrap Fusion

ORGANISM(S): Streptococcus Pneumoniae Tigr4

SUBMITTER: Jens Hör

LAB HEAD: Jörg Vogel

PROVIDER: PXD015835 | Pride | 2020-04-08

REPOSITORIES: Pride

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			Action	DRS
	2017-05-22_Fusion_11148.raw	Raw
	2017-05-22_Fusion_11149.raw	Raw
	2017-05-22_Fusion_11150.raw	Raw
	2017-05-22_Fusion_11151.raw	Raw
	2017-05-22_Fusion_11152.raw	Raw

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Publications

Grad-seq in a Gram-positive bacterium reveals exonucleolytic sRNA activation in competence control.

Hör Jens J Garriss Geneviève G Di Giorgio Silvia S Hack Lisa-Marie LM Vanselow Jens T JT Förstner Konrad U KU Schlosser Andreas A Henriques-Normark Birgitta B Vogel Jörg J

The EMBO journal 20200330 9

RNA-protein interactions are the crucial basis for many steps of bacterial gene expression, including post-transcriptional control by small regulatory RNAs (sRNAs). In stark contrast to recent progress in the analysis of Gram-negative bacteria, knowledge about RNA-protein complexes in Gram-positive species remains scarce. Here, we used the Grad-seq approach to draft a comprehensive landscape of such complexes in Streptococcus pneumoniae, in total determining the sedimentation profiles of ~ 88% o ...[more]

PMID: 32227509

Similar Datasets

A census of RNA/protein complexes in a model Gram-positive bacterium reveals exonuclease-mediated sRNA activation in competence regulation

Project description:Affinity purification samples performed with different sRNAs as the bait. Single lanes were cut from a gel and submitted for MS.

2020-04-08 | PXD015842 | Pride

Exonuclease-mediated sRNA activation in competence regulation in a Gram-positive bacterium

Project description:This SuperSeries is composed of the SubSeries listed below.

2020-03-20 | GSE145605 | GEO

A census of RNA/protein complexes in a model Gram-positive bacterium reveals exonuclease-mediated sRNA activation in competence regulation

Project description:RNA-protein interactions crucially underlie many steps of bacterial gene expression including post-transcriptional control by small regulatory RNAs (sRNAs). In stark contrast with recent progress in Gram-negative bacteria, knowledge about RNA and protein complexes in Gram-positive species remains scarce. Here, we used Grad-seq to draft a landscape of such complexes in Streptococcus pneumoniae, determining the sedimentation profiles of ~88% of the transcripts and ~62% of the proteins of this important human pathogen. Analysis of in-gradient distributions and subsequent tag-based protein capture identified interactions of the exoribonuclease Cbf1 (a.k.a. YhaM) with sRNAs that control bacterial competence. Contrary to expectation, the nucleolytic activity of Cbf1 stabilized these sRNAs, thereby promoting their function as repressors of competence. These results illustrate how this first RNA/protein complexome resource for a Gram-positive species can be utilized to identify new molecular factors in RNA-based regulation of pathways with relevance to bacterial virulence.

2021-01-27 | GSE138732 | GEO

Grad-seq in a Gram-positive bacterium reveals exonucleolytic sRNA activation in competence control.

Project description:RNA-protein interactions are the crucial basis for many steps of bacterial gene expression, including post-transcriptional control by small regulatory RNAs (sRNAs). In stark contrast to recent progress in the analysis of Gram-negative bacteria, knowledge about RNA-protein complexes in Gram-positive species remains scarce. Here, we used the Grad-seq approach to draft a comprehensive landscape of such complexes in Streptococcus pneumoniae, in total determining the sedimentation profiles of ~ 88% of the transcripts and ~ 62% of the proteins of this important human pathogen. Analysis of in-gradient distributions and subsequent tag-based protein capture identified interactions of the exoribonuclease Cbf1/YhaM with sRNAs that control bacterial competence for DNA uptake. Unexpectedly, the nucleolytic activity of Cbf1 stabilizes these sRNAs, thereby promoting their function as repressors of competence. Overall, these results provide the first RNA/protein complexome resource of a Gram-positive species and illustrate how this can be utilized to identify new molecular factors with functions in RNA-based regulation of virulence-relevant pathways.

| S-EPMC7196914 | biostudies-literature

Grad-seq in a Gram-positive bacterium reveals exonucleolytic sRNA activation in competence control

2020-03-20 | GSE145604 | GEO

Exonuclease-mediated sRNA activation in competence regulation in a Gram-positive bacterium

Project description:Exonuclease-mediated sRNA activation in competence regulation in a Gram-positive bacterium

| PRJNA607724 | ENA

A census of RNA/protein complexes in a model Gram-positive bacterium reveals exonuclease-mediated sRNA activation in competence regulation

Project description:A census of RNA/protein complexes in a model Gram-positive bacterium reveals exonuclease-mediated sRNA activation in competence regulation

| PRJNA576919 | ENA

Grad-seq shines light on unrecognized RNA and protein complexes in the model bacterium Escherichia coli

Project description:22 total proteome samples from E. coli wild type resolved on a glycerol gradient resulting in 20 fractions and a pellet fraction. A lysate sample is given as input control. Further, 2 affinity purification samples comparing YggL-3xFLAG pulldown vs. the wild type is given. In this case, each sample was run on a gel and the lanes cut into 15 gel pieces each.

2020-08-10 | PXD019900 | Pride

Grad-seq in a Gram-positive bacterium reveals exonucleolytic sRNA activation in competence control

Project description:Grad-seq in a Gram-positive bacterium reveals exonucleolytic sRNA activation in competence control

| PRJNA607729 | ENA

Localization pattern of conjugation machinery in a Gram-positive bacterium.

Project description:Conjugation is an efficient way for transfer of genetic information between bacteria, even between highly diverged species, and a major cause for the spreading of resistance genes. We have investigated the subcellular localization of several conserved conjugation proteins carried on plasmid pLS20 found in Bacillus subtilis. We show that VirB1, VirB4, VirB11, VirD2, and VirD4 homologs assemble at a single cell pole, but also at other sites along the cell membrane, in cells during the lag phase of growth. Bimolecular fluorescence complementation analyses showed that VirB4 and VirD4 interact at the cell pole and, less frequently, at other sites along the membrane. VirB1 and VirB11 also colocalized at the cell pole. Total internal reflection fluorescence microscopy showed that pLS20 is largely membrane associated and is frequently found at the cell pole, indicating that transfer takes place at the pole, which is a preferred site for the assembly of the active conjugation apparatus, but not the sole site. VirD2, VirB4, and VirD4 started to localize to the pole or the membrane in stationary-phase cells, and VirB1 and VirB11 were observed as foci in cells resuspended in fresh medium but no longer in cells that had entered exponential growth, although at least VirB4 was still expressed. These data reveal an unusual assembly/disassembly timing for the pLS20 conjugation machinery and suggest that specific localization of conjugation proteins in lag-phase cells and delocalization during growth are the reasons why pLS20 conjugation occurs only during early exponential phase.

| S-EPMC3209232 | biostudies-literature

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