Structure of the OMEGA nickase IsrB in complex with ωRNA and target DNA

Seiichi Hirano, Kalli Kappel, Han Altae-Tran, Guilhem Faure, Max E. Wilkinson, Soumya Kannan, F. Esra Demircioglu, Rui Yan, Momoko Shiozaki, Zhiheng Yu, Kira S. Makarova, Eugene V. Koonin, Rhiannon K. Macrae and Feng Zhang ()
Additional contact information
Seiichi Hirano: Broad Institute of MIT and Harvard
Kalli Kappel: Broad Institute of MIT and Harvard
Han Altae-Tran: Broad Institute of MIT and Harvard
Guilhem Faure: Broad Institute of MIT and Harvard
Max E. Wilkinson: Broad Institute of MIT and Harvard
Soumya Kannan: Broad Institute of MIT and Harvard
F. Esra Demircioglu: Broad Institute of MIT and Harvard
Rui Yan: Howard Hughes Medical Institute, Janelia Research Campus
Momoko Shiozaki: Howard Hughes Medical Institute, Janelia Research Campus
Zhiheng Yu: Howard Hughes Medical Institute, Janelia Research Campus
Kira S. Makarova: National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health
Eugene V. Koonin: National Center for Biotechnology Information, National Library of Medicine, National Institutes of Health
Rhiannon K. Macrae: Broad Institute of MIT and Harvard
Feng Zhang: Broad Institute of MIT and Harvard

Nature, 2022, vol. 610, issue 7932, 575-581

Abstract: Abstract RNA-guided systems, such as CRISPR–Cas, combine programmable substrate recognition with enzymatic function, a combination that has been used advantageously to develop powerful molecular technologies1,2. Structural studies of these systems have illuminated how the RNA and protein jointly recognize and cleave their substrates, guiding rational engineering for further technology development3. Recent work identified a new class of RNA-guided systems, termed OMEGA, which include IscB, the likely ancestor of Cas9, and the nickase IsrB, a homologue of IscB lacking the HNH nuclease domain4. IsrB consists of only around 350 amino acids, but its small size is counterbalanced by a relatively large RNA guide (roughly 300-nt ωRNA). Here, we report the cryogenic-electron microscopy structure of Desulfovirgula thermocuniculi IsrB (DtIsrB) in complex with its cognate ωRNA and a target DNA. We find the overall structure of the IsrB protein shares a common scaffold with Cas9. In contrast to Cas9, however, which uses a recognition (REC) lobe to facilitate target selection, IsrB relies on its ωRNA, part of which forms an intricate ternary structure positioned analogously to REC. Structural analyses of IsrB and its ωRNA as well as comparisons to other RNA-guided systems highlight the functional interplay between protein and RNA, advancing our understanding of the biology and evolution of these diverse systems.

Date: 2022
References: Add references at CitEc
Citations:

Downloads: (external link)
https://www.nature.com/articles/s41586-022-05324-6 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:610:y:2022:i:7932:d:10.1038_s41586-022-05324-6

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

DOI: 10.1038/s41586-022-05324-6

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 ().