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RNA processing by the CRISPR-associated NYN ribonuclease.

Haotian Chi1, Malcolm F White1

  • 1School of Biology, University of St Andrews, St Andrews, Fife KY16 9ST, U.K.

The Biochemical Journal
|May 24, 2024
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Summary

This study reveals that the NYN ribonuclease in the Bacteroides fragilis type III CRISPR system trims RNA intermediates into mature CRISPR RNAs (crRNAs) for effective bacterial antiviral defense.

Keywords:
CRISPRbacteriophagesendonucleasesimmunologynon-coding RNA

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Area of Science:

  • Microbiology
  • Molecular Biology
  • Immunology

Background:

  • CRISPR-Cas systems provide adaptive immunity in prokaryotes by targeting invasive nucleic acids.
  • Type III CRISPR systems utilize a Cas10 subunit to generate signaling molecules upon foreign RNA detection, activating effector proteins.
  • The Bacteroides fragilis type III CRISPR system produces SAM-AMP, which regulates a CorA-family effector for immunity.

Purpose of the Study:

  • To investigate the role of the CRISPR-associated NYN ribonuclease in the Bacteroides fragilis type III CRISPR system.
  • To determine the function of NYN in the maturation of CRISPR RNAs (crRNAs).

Main Methods:

  • Structural modeling of the NYN protein.
  • In vitro ribonuclease assays to assess enzyme activity.
  • Analysis of sequence-nonspecific ssRNA cleavage.

Main Results:

  • NYN exhibits sequence-nonspecific, Mn2+-dependent single-stranded RNA (ssRNA) cleavage activity.
  • Structural modeling provided insights into NYN's potential function.
  • The findings suggest NYN is involved in processing crRNA intermediates.

Conclusions:

  • The NYN ribonuclease likely plays a crucial role in trimming crRNA intermediates to mature crRNAs for type III CRISPR antiviral defense.
  • This research elucidates the function of CRISPR-associated NYN proteins in bacterial immunity.
  • The study highlights the intricate mechanisms of bacterial CRISPR-mediated defense strategies.