Selective TnsC recruitment enhances the fidelity of RNA-guided transposition

Florian T. Hoffmann, Minjoo Kim, Leslie Y. Beh, Jing Wang, Phuc Leo H. Vo, Diego R. Gelsinger, Jerrin Thomas George, Christopher Acree, Jason T. Mohabir, Israel S. Fernández () and Samuel H. Sternberg ()
Additional contact information
Florian T. Hoffmann: Columbia University
Minjoo Kim: Columbia University
Leslie Y. Beh: Columbia University
Jing Wang: Columbia University
Phuc Leo H. Vo: Columbia University
Diego R. Gelsinger: Columbia University
Jerrin Thomas George: Columbia University
Christopher Acree: Columbia University
Jason T. Mohabir: Columbia University
Israel S. Fernández: Columbia University
Samuel H. Sternberg: Columbia University

Nature, 2022, vol. 609, issue 7926, 384-393

Abstract: Abstract Bacterial transposons are pervasive mobile genetic elements that use distinct DNA-binding proteins for horizontal transmission. For example, Escherichia coli Tn7 homes to a specific attachment site using TnsD1, whereas CRISPR-associated transposons use type I or type V Cas effectors to insert downstream of target sites specified by guide RNAs2,3. Despite this targeting diversity, transposition invariably requires TnsB, a DDE-family transposase that catalyses DNA excision and insertion, and TnsC, a AAA+ ATPase that is thought to communicate between transposase and targeting proteins4. How TnsC mediates this communication and thereby regulates transposition fidelity has remained unclear. Here we use chromatin immunoprecipitation with sequencing to monitor in vivo formation of the type I-F RNA-guided transpososome, enabling us to resolve distinct protein recruitment events before integration. DNA targeting by the TniQ–Cascade complex is surprisingly promiscuous—hundreds of genomic off-target sites are sampled, but only a subset of those sites is licensed for TnsC and TnsB recruitment, revealing a crucial proofreading checkpoint. To advance the mechanistic understanding of interactions responsible for transpososome assembly, we determined structures of TnsC using cryogenic electron microscopy and found that ATP binding drives the formation of heptameric rings that thread DNA through the central pore, thereby positioning the substrate for downstream integration. Collectively, our results highlight the molecular specificity imparted by consecutive factor binding to genomic target sites during RNA-guided transposition, and provide a structural roadmap to guide future engineering efforts.

Date: 2022
References: Add references at CitEc
Citations: View citations in EconPapers (1)

Downloads: (external link)
https://www.nature.com/articles/s41586-022-05059-4 Abstract (text/html)
Access to the full text of the articles in this series is restricted.

Related works:
This item may be available elsewhere in EconPapers: Search for items with the same title.

Export reference: BibTeX RIS (EndNote, ProCite, RefMan) HTML/Text

Persistent link: https://EconPapers.repec.org/RePEc:nat:nature:v:609:y:2022:i:7926:d:10.1038_s41586-022-05059-4

Ordering information: This journal article can be ordered from
https://www.nature.com/

DOI: 10.1038/s41586-022-05059-4

Access Statistics for this article

Nature is currently edited by Magdalena Skipper

More articles in Nature from Nature
Bibliographic data for series maintained by Sonal Shukla () and Springer Nature Abstracting and Indexing ().