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Structures of the holo CRISPR RNA-guided transposon integration complex
Open accessPreprint — published as 10.1038/s41586-022-05573-5 · Paper page
Abstract
CRISPR-associated transposons (CAST) are programmable mobile elements that insert large DNA cargo by an RNA-guided mechanism. Multiple conserved components act in concert at the target site through formation of an integration complex (transpososome). We reconstituted the type V-K CAST transpososome from Scytonema hofmannii (ShCAST) and determined the structure using cryo-EM. Transpososome architecture ensures orientation-specific association: AAA+ regulator TnsC has defined polarity and length, with dedicated interaction interfaces for other CAST components. Interestingly, transposase (TnsB)-TnsC interactions we observe contribute to stimulating TnsC’s ATP hydrolysis activity. TnsC deviates from previously observed helical configurations of TnsC, and target DNA does not track with TnsC protomers. Consequently, TnsC makes new, functionally important protein-DNA interactions throughout the transpososome. Finally, two distinct transpososome populations suggests that associations with the CRISPR effector are flexible. These ShCAST transpososome structures significantly enhances our understanding of CAST transposition systems and suggests avenues for improving CAST transposition for precision genome-editing applications.
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Highly accurate protein structure prediction with AlphaFold
Jumper, J., Evans, R., Pritzel, A., Green, T., Figurnov, M., Ronneberger, O., Tunyasuvunakool, K., Bates, R., Žídek, A., Potapenko, A., et al. (2021) · Nature · doi:10.1038/s41586-021-03819-2 · Cited by 43,764
Flagged first, then by citations
Research works printed but not linked: 45
- 10.1073/pnas.1709035114 · DOI
- 10.1126/science.aax9181 · DOI
- 10.1038/s41586-019-1323-z · DOI
- 2019, CRISPR-Cas in mobile genetic elements: counter-defence and beyond, Nat Rev Microbiol, 17, 513, 10.1038/s41579-019-0204-7 · DOI
- 10.1016/j.cell.2020.11.005 · DOI
- M. Saito et al., Dual modes of CRISPR-associated transposon homing. Cell, (2021). · DOI
- 10.1128/mBio.02938-21 · DOI
- 10.1111/mmi.14383 · DOI
- 10.1038/s41586-019-1849-0 · DOI
- M. Schmitz , I. Querques , S. Oberli , C. Chanez , M. Jinek , Structural basis for RNA-mediated assembly of type V CRISPR-associated transposons. bioRxiv, (2022). · DOI
- 10.1093/emboj/17.19.5822 · DOI
- F. T. Hoffmann et al., Selective TnsC recruitment enhances the fidelity of RNA-guided transposition. Nature, (2022). · DOI
- 10.1073/pnas.1409869111 · DOI
- 10.1101/gad.2.2.137 · DOI
- 2022, Evolutionary and mechanistic diversity of Type I-F CRISPR-associated transposons, Mol Cell, 82, 616, 10.1016/j.molcel.2021.12.021 · DOI
- I. Querques , M. Schmitz , S. Oberli , C. Chanez , M. Jinek , Target site selection and remodelling by type V CRISPR-transposon systems. Nature, (2021). · DOI
- R. Xiao et al., Structural basis of target DNA recognition by CRISPR-Cas12k for RNA-guided DNA transposition. Mol Cell, (2021). · DOI
- J. U. Park , A. W. Tsai , T. H. Chen , J. E. Peters , E. H. Kellogg , Mechanistic details of CRISPR-associated transposon recruitment and integration revealed by cryo-EM. Proc Natl Acad Sci U S A 119, e2202590119 (2022). · DOI
- 10.1126/science.abi8976 · DOI
- 10.1038/s41594-022-00724-8 · DOI
- 10.1006/jmbi.1996.0164 · DOI
- 10.1038/nature11602 · DOI · Paper page
- 10.2144/000113751 · DOI
- 10.1073/pnas.2009707117 · DOI
- A. Patel , D. Toso , A. Litvak , E. Nogales , Efficient graphene oxide coating improves cryo-EM sample preparation and data collection from tilted grids. bioRxiv, (2021). · DOI
- 10.1016/j.jsb.2005.03.010 · DOI
- 10.1038/nmeth.4193 · DOI
- 10.1016/j.jsb.2009.01.002 · DOI
- 10.1038/nmeth.4169 · DOI · Paper page
- 10.1016/j.jsb.2012.09.006 · DOI · Paper page
- 10.1016/bs.mie.2016.04.012 · DOI
- J. Zivanov et al., New tools for automated high-resolution cryo-EM structure determination in RELION-3. Elife 7, (2018). · DOI
- 10.1107/S205225251801463X · DOI
- 10.1002/jcc.20084 · DOI
- A. Punjani , D. J. Fleet , 3D variability analysis: Resolving continuous flexibility and discrete heterogeneity from single particle cryo-EM. J Struct Biol 213, 107702 (2021). · DOI
- 10.1107/S0907444910007493 · DOI · Paper page
- 10.1107/S2059798318009324 · DOI
- 10.1002/pro.3235 · DOI · Paper page
- 10.1002/pro.3330 · DOI · Paper page
- 10.1016/j.jsb.2005.07.007 · DOI
- 2019, Real-time cryo-electron microscopy data preprocessing with Warp, Nat Methods, 16, 1146, 10.1038/s41592-019-0580-y · DOI
- 2019, Positive-unlabeled convolutional neural networks for particle picking in cryo-electron micrographs, Nat Methods, 16, 1153, 10.1038/s41592-019-0575-8 · DOI
- 2021, Local computational methods to improve the interpretability and analysis of cryo-EM maps, Nat Commun, 12, 1240, 10.1038/s41467-021-21509-5 · DOI
- W. M. Cianfrocco MA , Youn C , Wagner R , Leschziner AE. , COSMIC²: A Science Gateway for Cryo-Electron Microscopy Structure Determination. Practice & Experience in Advanced Research Computing, (2017). · DOI
- 10.1016/B978-0-12-381270-4.00019-6 · DOI
Metadata
- Authors
- Park, J.-U., Tsai, A. W.-L., Rizo, A. N., Truong, V. H., Wellner, T. X., Schargel, R. D., & Kellogg, E. H.
- Affiliations
- Cornell University (US)
- Corresponding author
- Kellogg, Elizabeth H.
- ORCID
- Venue
- Nature
- Publisher
- openRxiv
- Published
- 2022-10-12
- Volume
- 613
- Issue
- 7945
- Pages
- 775-782
- ISSN
- 0028-0836 (ISSN-L)
- PMC
- PMC9876797
- OpenAlex
- W4310155672
- Cited by
- 80
- References
- 1
- Citations by year
- 2026: 22 · 2025: 21 · 2024: 21 · 2023: 15 · 2022: 1
- FWCI
- 4.73
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- cc-by
- Locations
- Nature (publishedVersion, cc-by)
- bioRxiv (Cold Spring Harbor Laboratory) (acceptedVersion)
- PubMed Central (publishedVersion, cc-by)
- PubMed (publishedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- Funders
- Simons Foundation (SF 349247)
- National Institutes of Health (1s10od030470-01, MCB200090)
- National Institute of General Medical Sciences (U24GM129539, R01GM144566)
- Common Fund (U24 GM129539)
- 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 › Biochemical and Structural Characterization
- Keywords
- transpososome, cascades, ribosomal protein S15, R-loops, protein-DNA interactions, transposition mechanism, genome editing
- MeSH terms
- Gene Editing, RNA, Guide, CRISPR-Cas Systems, Cyanobacteria, Bacterial Proteins, DNA Transposable Elements, DNA-Binding Proteins, Endodeoxyribonucleases, Transposases, Holoenzymes, Cryoelectron Microscopy, Multiprotein Complexes, Ribosome Subunits, Clustered Regularly Interspaced Short Palindromic Repeats, CRISPR-Cas Systems, CRISPR-Associated Proteins
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