Accommodation of helical imperfections in rhodobacter sphaeroides argonaute ternary complexes with guide RNA and target DNA Journal Article

   

Authors:	Liu, Y.; Esyunina, D.; Olovnikov, I.; Teplova, M.; Kulbachinskiy, A.; Aravin, A. A.; Patel, D. J.
Article Title:	Accommodation of helical imperfections in rhodobacter sphaeroides argonaute ternary complexes with guide RNA and target DNA
Abstract:	Prokaryotic Argonaute (Ago) proteins were recently shown to target foreign genetic elements, thus making them a perfect model for studies of interference mechanisms. Here, we study interactions of Rhodobacter sphaeroides Ago (RsAgo) with guide RNA (gRNA) and fully complementary or imperfect target DNA (tDNA) using biochemical and structural approaches. We show that RsAgo can specifically recognize both the first nucleotide in gRNA and complementary nucleotide in tDNA, and both interactions contribute to nucleic acid binding. Non-canonical pairs and bulges on the target strand can be accommodated by RsAgo with minimal perturbation of the duplex but significantly reduce RsAgo affinity to tDNA. Surprisingly, mismatches between gRNA and tDNA induce dissociation of the guide-target duplex from RsAgo. Our results reveal plasticity in the ability of Ago proteins to accommodate helical imperfections, show how this might affect the efficiency of RNA silencing, and suggest a potential mechanism for guide release and Ago recycling. © 2018 The Authors Prokaryotic Argonaute proteins (pAgos) target foreign genetic elements. Liu et al. structurally and functionally characterize interactions of pAgo with guide RNA and fully complementary or imperfectly targeted DNA, reveal plasticity in the ability of Ago proteins to accommodate helical imperfections, and suggest a potential mechanism for guide release.
Keywords:	guide rna; non-canonical base pairs and bulges; rhodobacter sphaeroides argonaute; rna-dna heteroduplex; rsago; target dna
Journal Title:	Cell Reports
Volume:	24
Issue:	2
ISSN:	2211-1247
Publisher:	Cell Press
Date Published:	2018-07-10
Start Page:	453
End Page:	462
Language:	English
DOI:	10.1016/j.celrep.2018.06.021
PROVIDER:	scopus
PUBMED:	29996105
PMCID:	PMC6269105
DOI/URL:	https://www.scopus.com/inward/record.uri?eid=2-s2.0-85049353079&doi=10.1016%2fj.celrep.2018.06.021&partnerID=40&md5=11698b8721e146f26e5549878837f307
Notes:	Article -- Export Date: 1 August 2018 -- Source: Scopus

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