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RNA-triggered protein cleavage and cell growth arrest by the type III-E CRISPR nuclease-protease

Kazuki Kato1, Sae Okazaki1, Cian Schmitt-Ulms2

  • 1Structural Biology Division, Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.

Science (New York, N.Y.)
|November 24, 2022
PubMed

Insights

The CRISPR-Cas7-11 system uses a CRISPR RNA (crRNA) to guide RNA cleavage and protein degradation. This RNA-dependent nuclease-protease complex can be engineered for programmable RNA sensing.

Area of Science:

  • Molecular Biology
  • Microbiology
  • Biochemistry

Background:

  • Type III-E CRISPR-Cas7-11 is an RNA-guided effector complex.
  • It involves the protease Csx29 and accessory proteins Csx30 and Csx31.

Purpose of the Study:

  • To elucidate the structural and functional mechanisms of the Cas7-11-crRNA-Csx29 complex.
  • To investigate the role of Csx30 cleavage in cellular processes and engineer the system for programmable RNA sensing.

Main Methods:

  • Cryo-electron microscopy to determine complex structures.
  • Biochemical assays to study RNA cleavage and protein degradation.
  • Bacterial reconstitution experiments to assess cellular effects and engineering in mammalian cells.

Main Results:

  • Cryo-EM structures revealed target RNA-induced conformational changes in Csx29.
  • Csx29 cleaves Csx30 in a target RNA-dependent manner, producing toxic fragments that arrest bacterial growth.
  • Csx30-Csx31-RpoE interaction suggests a role in modulating cellular responses to infection.
  • The system was engineered for programmable RNA sensing in mammalian cells.

Conclusions:

  • The Cas7-11-Csx29 complex functions as an RNA-dependent nuclease-protease.
  • This system offers a versatile platform for RNA-guided gene regulation and sensing applications.

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