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MS2-Affinity Purification Coupled with RNA Sequencing in Gram-Positive Bacteria
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Published on: February 23, 2021

SR1--a small RNA with two remarkably conserved functions.

Matthias Gimpel1, Heike Preis, Emanuel Barth

  • 1AG Bakteriengenetik, Lehrstuhl für Mikrobiologie und Mikrobengenetik, Friedrich-Schiller-Universität Jena, Philosophenweg 12, Jena D-07743, Germany.

Nucleic Acids Research
|October 5, 2012
PubMed
Summary
This summary is machine-generated.

SR1, a dual-function small regulatory RNA (sRNA), regulates gene expression and stabilizes mRNA by binding to GapA protein. Its functions and structure are highly conserved across Bacillus species, indicating ancient evolutionary importance.

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Area of Science:

  • * Molecular Biology
  • * RNA Regulation
  • * Microbial Genetics

Background:

  • * SR1 is a unique small regulatory RNA (sRNA) with dual functions: base-pairing regulation of ahrC mRNA and peptide encoding of SR1P.
  • * SR1P peptide stabilizes gapA mRNA, crucial for glycolytic conditions.
  • * SR1 transcription is repressed by CcpN and CcpA under specific conditions.

Purpose of the Study:

  • * To investigate the evolutionary conservation and functional integrity of SR1 and its encoded peptide (SR1P) across various Bacillus species.
  • * To explore the regulatory mechanisms controlling SR1 expression, including CcpN and CcpA involvement.
  • * To assess the functional complementation and binding capabilities of SR1 homologues in Bacillus subtilis.

Main Methods:

  • * Bioinformatic analysis to identify SR1 homologues in Bacillus, Geobacillus, Anoxybacillus, and Brevibacillus species.
  • * Construction and analysis of transcriptional lacZ fusions to study SR1 expression regulation.
  • * Functional complementation assays in a Bacillus subtilis Δsr1 strain to evaluate SR1P's ability to stabilize gapA mRNA.
  • * In vitro binding assays to assess the base-pairing function of SR1/ahrC interactions.

Main Results:

  • * 23 SR1 homologues were identified, exhibiting high structural and sequence similarity to Bacillus subtilis SR1.
  • * SR1 expression is primarily regulated by CcpN and glucose-sensitive, with some homologues showing additional repression.
  • * Most tested SR1P homologues could complement the B. subtilis Δsr1 strain, but only half tightly bound GapA.
  • * In vitro assays confirmed the conserved base-pairing function of SR1/ahrC interactions despite sequence divergence.

Conclusions:

  • * SR1 represents a highly conserved ancient regulatory system with dual functions, preserved for over a billion years.
  • * Both the RNA-based regulatory function and the peptide-mediated mRNA stabilization are evolutionarily maintained.
  • * SR1 and SR1P play a conserved role in regulating gene expression within the Bacillus genus.