8jpx

Structural highlights

Function

AGO_PYRFU A DNA-guided ssDNA endonuclease that may play a role in defense against invading mobile genetic elements. Uses short 5'-phospho-ssDNA sequences as guides (gDNA) to bind complementary target strands, resulting in cleavage of the target DNA (tDNA). Endonucleolytically cleaves DNA in short dsDNA (the gDNA indicates where to cleave on the tDNA). Efficient guide-dependent target DNA cleavage requires a minimal gDNA length of 15 nucleotides (nt) and works up to at least 31 nt. Overexpression decreases plasmid transformation efficiency. Has no appreciable activity with gRNA or on target RNA. Also has guide-independent activity on plasmid DNA called 'chopping' (PubMed:25925567). The cleavage site is 10 nucleotides (nt) downstream of the target residue base-paired with the 5'-end of the gDNA, cleavage is insensitive to adenine methylation. DNA cleavage produces 5'-phosphomonoesters (as it can be ligated by T4 DNA ligase) (PubMed:28165224).^{[1] [2]}

References

1. ↑ Swarts DC, Hegge JW, Hinojo I, Shiimori M, Ellis MA, Dumrongkulraksa J, Terns RM, Terns MP, van der Oost J. Argonaute of the archaeon Pyrococcus furiosus is a DNA-guided nuclease that targets cognate DNA. Nucleic Acids Res. 2015 May 26;43(10):5120-9. PMID:25925567 doi:10.1093/nar/gkv415
2. ↑ Enghiad B, Zhao H. Programmable DNA-Guided Artificial Restriction Enzymes. ACS Synth Biol. 2017 May 19;6(5):752-757. PMID:28165224 doi:10.1021/acssynbio.6b00324