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Grad-seq in a Gram-positive bacterium reveals exonucleolytic sRNA activation in competence control
Jens Hör1, Geneviève Garriss2, Silvia Di Giorgio1,3
1Institute of Molecular Infection Biology, University of Würzburg, Würzburg, Germany.
The EMBO Journal
|April 1, 2020
Summary
Researchers mapped RNA-protein complexes in Streptococcus pneumoniae, revealing how the Cbf1 exoribonuclease stabilizes small regulatory RNAs (sRNAs) to control DNA uptake and bacterial competence.
Area of Science:
- Bacteriology
- Molecular Biology
- Genetics
Background:
- RNA-protein interactions regulate bacterial gene expression, particularly post-transcriptional control by small regulatory RNAs (sRNAs).
- Understanding of RNA-protein complexes is limited in Gram-positive bacteria compared to Gram-negative species.
Purpose of the Study:
- To comprehensively map RNA-protein complexes in the Gram-positive bacterium Streptococcus pneumoniae.
- To identify novel RNA-binding proteins and their functions in gene regulation.
Main Methods:
- Utilized the Grad-seq approach to determine sedimentation profiles of transcripts and proteins.
- Employed tag-based protein capture to identify RNA-protein interactions.
Main Results:
- Generated a comprehensive RNA/protein complexome map for Streptococcus pneumoniae, covering approximately 88% of transcripts and 62% of proteins.
- Identified interactions between the exoribonuclease Cbf1/YhaM and sRNAs involved in regulating competence for DNA uptake.
- Discovered that Cbf1's nucleolytic activity unexpectedly stabilizes these sRNAs, enhancing their role as competence repressors.
Conclusions:
- This study provides the first RNA/protein complexome resource for a Gram-positive bacterium.
- Demonstrates the utility of this resource for discovering new factors in RNA-based regulation of virulence pathways.
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