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PreprintCited by 24CRISPR and Genetic EngineeringBacterial Genetics and BiotechnologyRNA regulation and disease

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Structure and mechanism of the type I-G CRISPR effector

Open access
Shangguan, Qilin, Graham, Shirley, Sundaramoorthy, Ramasubramanian, White, Malcolm F·2022·Nucleic Acids Research
DOI 10.1101/2022.08.08.503147PMC PMC9638904

Preprint — published as 10.1093/nar/gkac925 · Paper page

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Abstract

Type I CRISPR systems are the most common CRISPR type found in bacteria. They use a multisubunit effector, guided by crRNA, to detect and bind dsDNA targets, forming an R-loop and recruiting the Cas3 enzyme to facilitate target DNA destruction, thus providing immunity against mobile genetic elements. Subtypes have been classified into families A-G, with type I-G being the least well understood. Here, we report the composition, structure and function of the type I-G Cascade CRISPR effector from Thioalkalivibrio sulfidiphilus , revealing key new molecular details. The unique Csb2 subunit processes pre-crRNA, remaining bound to the 3’ end of the mature crRNA, and seven Cas7 subunits form the backbone of the effector. Cas3 associates stably with the effector complex via the Cas8g subunit and is important for target DNA recognition. Structural analysis by cryo-Electron Microscopy reveals a strikingly curved backbone conformation with Cas8g spanning the belly of the structure. Type I-G Cascade is one of the most streamlined Class 1 CRISPR effectors. These biochemical and structural insights shed new light on the diversity of type I systems and open the way to applications in genome engineering.

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Metadata

Authors
Shangguan, Q., Graham, S., Sundaramoorthy, R., & White, M. F.
Affiliations
  • University of St Andrews (GB)
  • University of Dundee (GB)
Corresponding author
  • Sundaramoorthy, Ramasubramanian
  • White, Malcolm F
ORCID
Venue
Nucleic Acids Research
Publisher
openRxiv
Published
2022-08-08
Volume
50
Issue
19
Pages
11214-11228
ISSN
0305-1048 (ISSN-L)
DOI
10.1101/2022.08.08.503147
PMC
PMC9638904
OpenAlex
W4307492594
Cited by
24
References
1
Citations by year
2026: 5 · 2025: 4 · 2024: 7 · 2023: 8
FWCI
1.55
Type
journal article
Language
en
Open access
Licence
cc-by
Locations
  • Nucleic Acids Research (publishedVersion, cc-by)
  • bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion, cc-by)
  • PubMed Central (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • Discovery Research Portal (University of Dundee) (publishedVersion, cc-by)
  • St Andrews Research Repository (St Andrews Research Repository) (submittedVersion, cc-by)
Funders
  • University of St Andrews
  • China Scholarship Council (202008060345)
  • Directorate for Biological Sciences
  • Medical Research Council (MR/S021647/1)
  • Biotechnology and Biological Sciences Research Council (BB/S000313/1)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › CRISPR and Genetic Engineering
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Genetics › Bacterial Genetics and Biotechnology
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › RNA regulation and disease
Keywords
Cascade complex, Cas3, R-loops, DNA cleavage, bacterial immunity, cryo-EM structure
MeSH terms
Bacterial Proteins, DNA, Cryoelectron Microscopy, Ectothiorhodospiraceae, CRISPR-Cas Systems, CRISPR-Associated Proteins

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