5ux0
From Proteopedia
(Difference between revisions)
| (4 intermediate revisions not shown.) | |||
| Line 1: | Line 1: | ||
| - | '''Unreleased structure''' | ||
| - | + | ==X-ray crystal structure of Marinitoga piezophila Argonaute in complex with 5' OH guide RNA and target DNA== | |
| + | <StructureSection load='5ux0' size='340' side='right'caption='[[5ux0]], [[Resolution|resolution]] 3.20Å' scene=''> | ||
| + | == Structural highlights == | ||
| + | <table><tr><td colspan='2'>[[5ux0]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Marinitoga_piezophila Marinitoga piezophila] and [https://en.wikipedia.org/wiki/Synthetic_construct Synthetic construct]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5UX0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=5UX0 FirstGlance]. <br> | ||
| + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 3.197Å</td></tr> | ||
| + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=5ux0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5ux0 OCA], [https://pdbe.org/5ux0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=5ux0 RCSB], [https://www.ebi.ac.uk/pdbsum/5ux0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=5ux0 ProSAT]</span></td></tr> | ||
| + | </table> | ||
| + | == Function == | ||
| + | [https://www.uniprot.org/uniprot/AGO_MARPK AGO_MARPK] An RNA-guided ssDNA endonuclease that may play a role in defense against invading mobile genetic elements. Uses short 5'-OH-ssRNA sequences as guides (gRNA) to bind complementary target DNA (tDNA) or target RNA resulting in target cleavage. The cleavage site is 10 nucleotides (nt) downstream of the target residue base-paired with the 5'-end of the gRNA. Reaction rates are fastest on 5'-OH-gRNA:tDNA followed by 5'-OH-gRNA:target RNA. gRNA between 17-21 nt supports equivalent rates of cleavage, has no preferred 5'-nt. Has weak activity on tDNA with 5'-phospho-gRNA, yielding products 1-2 nt longer (PubMed:27035975). Unlike other characterized prokaryotic Ago proteins symmetric mismatches centered around the cleavage site reduce cleavage efficiency (PubMed:28520746).<ref>PMID:27035975</ref> <ref>PMID:28520746</ref> | ||
| + | <div style="background-color:#fffaf0;"> | ||
| + | == Publication Abstract from PubMed == | ||
| + | Argonaute (Ago) proteins are widespread in prokaryotes and eukaryotes and share a four-domain architecture capable of RNA- or DNA-guided nucleic acid recognition. Previous studies identified a prokaryotic Argonaute protein from the eubacterium Marinitoga piezophila (MpAgo), which binds preferentially to 5'-hydroxylated guide RNAs and cleaves single-stranded RNA (ssRNA) and DNA (ssDNA) targets. Here we present a 3.2 A resolution crystal structure of MpAgo bound to a 21-nucleotide RNA guide and a complementary 21-nucleotide ssDNA substrate. Comparison of this ternary complex to other target-bound Argonaute structures reveals a unique orientation of the N-terminal domain, resulting in a straight helical axis of the entire RNA-DNA heteroduplex through the central cleft of the protein. Additionally, mismatches introduced into the heteroduplex reduce MpAgo cleavage efficiency with a symmetric profile centered around the middle of the helix. This pattern differs from the canonical mismatch tolerance of other Argonautes, which display decreased cleavage efficiency for substrates bearing sequence mismatches to the 5' region of the guide strand. This structural analysis of MpAgo bound to a hybrid helix advances our understanding of the diversity of target recognition mechanisms by Argonaute proteins. | ||
| - | + | DNA recognition by an RNA-guided bacterial Argonaute.,Doxzen KW, Doudna JA PLoS One. 2017 May 17;12(5):e0177097. doi: 10.1371/journal.pone.0177097., eCollection 2017. PMID:28520746<ref>PMID:28520746</ref> | |
| - | + | From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |
| - | [[Category: | + | </div> |
| - | [[Category: Doudna | + | <div class="pdbe-citations 5ux0" style="background-color:#fffaf0;"></div> |
| - | [[Category: Doxzen | + | |
| + | ==See Also== | ||
| + | *[[Argonaute 3D structures|Argonaute 3D structures]] | ||
| + | == References == | ||
| + | <references/> | ||
| + | __TOC__ | ||
| + | </StructureSection> | ||
| + | [[Category: Large Structures]] | ||
| + | [[Category: Marinitoga piezophila]] | ||
| + | [[Category: Synthetic construct]] | ||
| + | [[Category: Doudna JA]] | ||
| + | [[Category: Doxzen KW]] | ||
Current revision
X-ray crystal structure of Marinitoga piezophila Argonaute in complex with 5' OH guide RNA and target DNA
| |||||||||||
