PXD068253

PXD068253 is an original dataset announced via ProteomeXchange.

Title	Structure of the minimal type I-F2 CRISPR-Cas DNA-interference complex
Description	CRISPR–Cas systems provide adaptive immunity in prokaryotes by targeting and degrading invasive genetic elements. Among them, the type I-F2 system represents the most compact type I CRISPR–Cas system, distinguished by the absence of small and large subunits. These subunits are otherwise essential for target recognition and nuclease recruitment. To elucidate how this minimal system mediates DNA interference, we determined the cryo-electron microscopy (cryo-EM) structure of the I-F2 Cascade complex bound to its target DNA and the effector helicase-nuclease Cas3. Our structure reveals a unique mechanism of Cas3 recruitment, predominantly mediated by the Cas7 subunit. We show how the helicase and C-terminal domains of Cas3 engage the single-stranded DNA to initiate directional DNA-unwinding and degradation. These findings uncover key mechanistic adaptations that enable efficient interference in the absence of canonical subunits and emphasize the mechanistic diversity among closely related type I systems, including I-E, I-F1, and I-F2. These insights provide a structural basis for engineering the minimal I-F2 system for genome editing and biotechnological applications.
HostingRepository	PRIDE
AnnounceDate	2026-03-09
AnnouncementXML	Submission_2026-03-08_20:33:34.960.xml
DigitalObjectIdentifier
ReviewLevel	Peer-reviewed dataset
DatasetOrigin	Original dataset
RepositorySupport	Unsupported dataset by repository
PrimarySubmitter	Wieland Steinchen
SpeciesList	scientific name: Shewanella putrefaciens CN-32; NCBI TaxID: NEWT:319224;
ModificationList	No PTMs are included in the dataset
Instrument	Synapt MS

Revision	Datetime	Status	ChangeLog Entry
0	2025-09-10 04:45:40	ID requested
⏵ 1	2026-03-08 20:33:35	announced

Perry TN, Mais CN, Sanchez-Londono M, Steinchen W, Plitzko PA, Randau L, Pausch P, Innis CA, Bange G, Structural basis of Cas8-independent Cas3 recruitment in Type I-F2 CRISPR-Cas. Nucleic Acids Res, 54(5):(2026) [pubmed]

10.1093/nar/gkag136;

ProteomeXchange project tag: Hydrogen Deuterium Exchange (HDX-MS)

submitter keyword: cryo-EM, Shewanella putrefaciens, HDX-MS,DNA interference, interference complex

Gert Bange
contact affiliation	Philipps University Marburg, Center for Synthetic Microbiology & Department of Chemistry, 35043 Marburg, Germany
contact email	bangeg@staff.uni-marburg.de
lab head
Wieland Steinchen
contact affiliation	Philipps-University Marburg, Department of Chemistry & SYNMIKRO, Karl-von-Frisch-Straße 14, 35043 Marburg, GERMANY
contact email	wieland.steinchen@synmikro.uni-marburg.de
dataset submitter

Dataset FTP location NOTE: Most web browsers have now discontinued native support for FTP access within the browser window. But you can usually install another FTP app (we recommend FileZilla) and configure your browser to launch the external application when you click on this FTP link. Or otherwise, launch an app that supports FTP (like FileZilla) and use this address: ftp://ftp.pride.ebi.ac.uk/pride/data/archive/2026/03/PXD068253

PRIDE project URI

• PRIDE
  1. PXD068253
     1. Label: PRIDE project
     2. Name: Structure of the minimal type I-F2 CRISPR-Cas DNA-interference complex