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TitlePhage-associated Cas12p nucleases require binding to bacterial thioredoxin for activation and cleavage of target DNA.
Journal, issue, pagesNat Microbiol, Vol. 11, Issue 1, Page 81-93, Year 2026
Publish dateJan 2, 2026
AuthorsZhipeng Wang / Yujue Wang / Hui Gao / Jiani Dai / Na Tang / Yannan Wang / Quanjiang Ji /
PubMed AbstractThe evolutionary competition within phage-host systems led to the emergence of CRISPR-Cas defence mechanisms in bacteria and anti-CRISPR elements in bacteriophages. Although anti-CRISPR elements are ...The evolutionary competition within phage-host systems led to the emergence of CRISPR-Cas defence mechanisms in bacteria and anti-CRISPR elements in bacteriophages. Although anti-CRISPR elements are well characterized, the role of bacterial factors that influence CRISPR-Cas efficacy has been comparatively overlooked. Type V CRISPR-Cas12 systems display striking functional and mechanistic diversity for nucleic acid targeting. Here we use a bioinformatic approach to identify Cas12p, a phage-associated nuclease that forms complexes with the bacterial thioredoxin protein TrxA to enable target DNA degradation. This represents an unexpected phage-bacteria interaction, in which the bacteriophage co-opts a bacterial factor to augment its own genome degradation machinery, potentially against competing phages. Biochemical characterization, cryo-EM-based structural analysis of the Cas12p-TrxA-sgRNA-dsDNA complex at 2.67 Å and bacterial defence assays reveal that TrxA directly binds and activates Cas12p, enabling its nuclease activity and subsequent CRISPR immunity. These findings expand our understanding of the multilayered intricacies of phage-bacteria molecular interactions.
External linksNat Microbiol / PubMed:41492065
MethodsEM (single particle)
Resolution2.67 - 3.2 Å
Structure data

61438

9jfs

EMDB-61438, PDB-9jfs:
Structure of Cas12p-TrxA-guide RNA-target DNA complex(29nt TS and 11nt NTS)
Method: EM (single particle) / Resolution: 2.67 Å

61449

9jg3

EMDB-61449, PDB-9jg3:
Structure of Cas12p-TrxA-guide RNA-target DNA complex(33-bp dsDNA)
Method: EM (single particle) / Resolution: 3.2 Å

Source
  • unidentified (others)
  • escherichia coli bl21(de3) (bacteria)
  • synthetic construct (others)
KeywordsRNA BINDING PROTEIN/RNA/DNA / CRISPR-Cas / nuclease / Heterodimer / RNA BINDING PROTEIN-RNA-DNA complex

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