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==Cryo-EM structure of the PseTnsAB paired-end complex (left end) in the presence of Mg== | |||
<StructureSection load='30jw' size='340' side='right'caption='[[30jw]], [[Resolution|resolution]] 2.51Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[30jw]] is a 8 chain structure with sequence from [https://en.wikipedia.org/wiki/Pseudoalteromonas_sp._S983 Pseudoalteromonas sp. S983]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=30JW OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=30JW FirstGlance]. <br> | |||
</td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">Electron Microscopy, [[Resolution|Resolution]] 2.51Å</td></tr> | |||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=MG:MAGNESIUM+ION'>MG</scene></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=30jw FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=30jw OCA], [https://pdbe.org/30jw PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=30jw RCSB], [https://www.ebi.ac.uk/pdbsum/30jw PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=30jw ProSAT]</span></td></tr> | |||
</table> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
CRISPR-associated transposons (CASTs) are Tn7-like elements that have co-opted RNA-guided CRISPR effectors for targeted DNA insertion. CASTs have been adapted as genome editing tools for programmable, site-specific integration. Among them, the type I-F system from Ps e udoalteromonas ( Pse CAST) shows uniquely robust activity in human cells, yet its mechanistic basis remains poorly understood. Here, we present structural and biochemical analysis of the Pse CAST transposase TnsAB. Biochemical reconstitution of transposon DNA excision defines key characteristics of the transposition mechanism. Cryogenic electron microscopy (cryo-EM) structures of Pse TnsAB paired-end complexes reveal molecular determinants of transpososome assembly, transposon end recognition and cleavage. We validate these findings using biochemical and in vivo assays of structure-based transposase mutants, and provide mechanistic insights into the enhanced activity of a laboratory-evolved TnsAB variant. Together, our studies highlight molecular features underlying the efficiency of natural and engineered type I-F transposases and establish a mechanistic framework for their continued rational optimization. | |||
Transposon end recognition and excision mechanisms of type I-F CRISPR-associated transposases.,Walter M, Finocchio G, Oberli S, Hammerschmid IC, Lampe GD, Karan J, Swartjes T, Sternberg SH, Jinek M, Querques I bioRxiv [Preprint]. 2026 May 7:2026.05.05.722991. doi: , 10.64898/2026.05.05.722991. PMID:42146515<ref>PMID:42146515</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
[[Category: | </div> | ||
<div class="pdbe-citations 30jw" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Large Structures]] | |||
[[Category: Pseudoalteromonas sp. S983]] | |||
[[Category: Finocchio G]] | |||
[[Category: Jinek M]] | |||
[[Category: Oberli S]] | |||
Latest revision as of 06:42, 3 June 2026
Cryo-EM structure of the PseTnsAB paired-end complex (left end) in the presence of Mg
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