5I4A

X-ray crystal structure of Marinitoga piezophila Argonaute in complex with 5' OH guide RNA

Summary for 5I4A

• Entry DOI: 10.2210/pdb5i4a/pdb
• Descriptor: Argonaute protein, RNA (5'-R(*UP*AP*UP*AP*CP*AP*AP*CP*CP*UP*AP*CP*UP*U)-3') (3 entities in total)
• Functional Keywords: argonaute, 5' oh, guide rna, rna binding protein-rna complex, rna binding protein/rna
• Biological source: Marinitoga piezophila (strain DSM 14283 / JCM 11233 / KA3); More
• Total number of polymer chains: 4
• Total formula weight: 165458.32

Authors

Doxzen, K.W.,Kaya, E.,Knoll, K.R.,Wilson, R.C.,Strutt, S.C.,Kranzusch, P.J.,Doudna, J.A. (deposition date: 2016-02-11, release date: 2016-03-30, Last modification date: 2024-03-06)

Primary citation

Kaya, E.,Doxzen, K.W.,Knoll, K.R.,Wilson, R.C.,Strutt, S.C.,Kranzusch, P.J.,Doudna, J.A.
A bacterial Argonaute with noncanonical guide RNA specificity.
Proc.Natl.Acad.Sci.USA, 113:4057-4062, 2016

PubMed Abstract: Eukaryotic Argonaute proteins induce gene silencing by small RNA-guided recognition and cleavage of mRNA targets. Although structural similarities between human and prokaryotic Argonautes are consistent with shared mechanistic properties, sequence and structure-based alignments suggested that Argonautes encoded within CRISPR-cas [clustered regularly interspaced short palindromic repeats (CRISPR)-associated] bacterial immunity operons have divergent activities. We show here that the CRISPR-associated Marinitoga piezophila Argonaute (MpAgo) protein cleaves single-stranded target sequences using 5'-hydroxylated guide RNAs rather than the 5'-phosphorylated guides used by all known Argonautes. The 2.0-Å resolution crystal structure of an MpAgo-RNA complex reveals a guide strand binding site comprising residues that block 5' phosphate interactions. Using structure-based sequence alignment, we were able to identify other putative MpAgo-like proteins, all of which are encoded within CRISPR-cas loci. Taken together, our data suggest the evolution of an Argonaute subclass with noncanonical specificity for a 5'-hydroxylated guide.

PubMed: 27035975
DOI: 10.1073/pnas.1524385113

Experimental method

X-RAY DIFFRACTION (1.949 Å)