Eukaryotic RNA-guided endonucleases evolved from a unique clade of bacterial enzymes

Author:

Yoon Peter H12,Skopintsev Petr23,Shi Honglue24ORCID,Chen LinXing25,Adler Benjamin A23ORCID,Al-Shimary Muntathar12,Craig Rory J3,Loi Kenneth J12,DeTurk Evan C23,Li Zheng6,Amerasekera Jasmine7,Trinidad Marena24,Nisonoff Hunter8,Chen Kai12ORCID,Lahiri Arushi12,Boger Ron23,Jacobsen Steve69ORCID,Banfield Jillian F25,Doudna Jennifer A123410111213ORCID

Affiliation:

1. Department of Molecular and Cell Biology, University of California , Berkeley ; Berkeley , CA , USA

2. Innovative Genomics Institute; University of California , Berkeley , CA , USA

3. California Institute for Quantitative Biosciences (QB3), University of California , Berkeley , CA , USA

4. Howard Hughes Medical Institute, University of California , Berkeley ; Berkeley,  CA , USA

5. Department of Earth and Planetary Science, University of California , Berkeley , CA , USA

6. Department of Molecular, Cell and Developmental Biology, University of California , Los Angeles,  CA , USA

7. Department of Human Genetics, University of California , Los Angeles,  CA , USA

8. Center for Computational Biology, University of California , Berkeley ; Berkeley, CA,  USA

9. Howard Hughes Medical Institute, University of California , Los Angeles  CA , USA

10. Gladstone Institutes ; San Francisco , CA , USA

11. Gladstone-UCSF Institute of Genomic Immunology ; San Francisco , CA , USA

12. Molecular Biophysics and Integrated Bioimaging Division, Lawrence Berkeley National Laboratory ; Berkeley , CA , USA

13. Department of Chemistry, University of California , Berkeley ; Berkeley , CA , USA

Abstract

Abstract RNA-guided endonucleases form the crux of diverse biological processes and technologies, including adaptive immunity, transposition, and genome editing. Some of these enzymes are components of insertion sequences (IS) in the IS200/IS605 and IS607 transposon families. Both IS families encode a TnpA transposase and a TnpB nuclease, an RNA-guided enzyme ancestral to CRISPR-Cas12s. In eukaryotes, TnpB homologs occur as two distinct types, Fanzor1s and Fanzor2s. We analyzed the evolutionary relationships between prokaryotic TnpBs and eukaryotic Fanzors, which revealed that both Fanzor1s and Fanzor2s stem from a single lineage of IS607 TnpBs with unusual active site arrangement. The widespread nature of Fanzors implies that the properties of this particular lineage of IS607 TnpBs were particularly suited to adaptation in eukaryotes. Biochemical analysis of an IS607 TnpB and Fanzor1s revealed common strategies employed by TnpBs and Fanzors to co-evolve with their cognate transposases. Collectively, our results provide a new model of sequential evolution from IS607 TnpBs to Fanzor2s, and Fanzor2s to Fanzor1s that details how genes of prokaryotic origin evolve to give rise to new protein families in eukaryotes.

Funder

Somatic Cell Genome Editing Program of the Common Fund of the National Institutes of Health

Swiss National Science Foundation Mobility fellowship

The Jane Coffin Childs Fund for Medical Research

National Institute of Health

m-CAFEs Microbial Community Analysis & Functional Evaluation in Soils

Lawrence Berkeley National Laboratory

US Department of Energy, Office of Science, Office of Biological & Environmental Research

U.S. Department of Energy

Publisher

Oxford University Press (OUP)

Subject

Genetics

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