Crosstalk between transposase subunits during cleavage of the mariner transposon

Mariner transposition is a complex reaction that involves three recombination sites and six strand breaking and joining reactions. This requires precise spatial and temporal coordination between the different components to ensure a productive outcome and minimize genomic instability. We have investi...

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Main Authors: Claeys Bouuaert, Corentin, Walker, Neil, Liu, Danxu, Chalmers, Ronald
Format: Article
Published: Oxford University Press 2014
Subjects:
Online Access:https://eprints.nottingham.ac.uk/41466/
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author Claeys Bouuaert, Corentin
Walker, Neil
Liu, Danxu
Chalmers, Ronald
author_facet Claeys Bouuaert, Corentin
Walker, Neil
Liu, Danxu
Chalmers, Ronald
author_sort Claeys Bouuaert, Corentin
building Nottingham Research Data Repository
collection Online Access
description Mariner transposition is a complex reaction that involves three recombination sites and six strand breaking and joining reactions. This requires precise spatial and temporal coordination between the different components to ensure a productive outcome and minimize genomic instability. We have investigated how the cleavage events are orchestrated within the mariner transpososome. We find that cleavage of the non-transferred strand is completed at both transposon ends before the transferred strand is cleaved at either end. By introducing transposon-end mutations that interfere with cleavage, but leave transpososome assembly unaffected, we demonstrate that a structural transition preceding transferred strand cleavage is coordinated between the two halves of the transpososome. Since mariner lacks the DNA hairpin intermediate, this transition probably reflects a reorganization of the transpososome to allow the access of different monomers onto the second pair of strands, or the relocation of the DNA within the same active site between two successive hydrolysis events. Communication between transposase subunits also provides a failsafe mechanism that restricts the generation of potentially deleterious double-strand breaks at isolated sites. Finally, we identify transposase mutants that reveal that the conserved WVPHEL motif provides a structural determinant of the coordination mechanism.
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spelling nottingham-414662020-05-04T16:48:04Z https://eprints.nottingham.ac.uk/41466/ Crosstalk between transposase subunits during cleavage of the mariner transposon Claeys Bouuaert, Corentin Walker, Neil Liu, Danxu Chalmers, Ronald Mariner transposition is a complex reaction that involves three recombination sites and six strand breaking and joining reactions. This requires precise spatial and temporal coordination between the different components to ensure a productive outcome and minimize genomic instability. We have investigated how the cleavage events are orchestrated within the mariner transpososome. We find that cleavage of the non-transferred strand is completed at both transposon ends before the transferred strand is cleaved at either end. By introducing transposon-end mutations that interfere with cleavage, but leave transpososome assembly unaffected, we demonstrate that a structural transition preceding transferred strand cleavage is coordinated between the two halves of the transpososome. Since mariner lacks the DNA hairpin intermediate, this transition probably reflects a reorganization of the transpososome to allow the access of different monomers onto the second pair of strands, or the relocation of the DNA within the same active site between two successive hydrolysis events. Communication between transposase subunits also provides a failsafe mechanism that restricts the generation of potentially deleterious double-strand breaks at isolated sites. Finally, we identify transposase mutants that reveal that the conserved WVPHEL motif provides a structural determinant of the coordination mechanism. Oxford University Press 2014-05-14 Article PeerReviewed Claeys Bouuaert, Corentin, Walker, Neil, Liu, Danxu and Chalmers, Ronald (2014) Crosstalk between transposase subunits during cleavage of the mariner transposon. Nucleic Acids Research, 42 (9). pp. 5799-5808. ISSN 1362-4962 mutation; dna; transposase; coordination; cytokinesis; dna transposons; double-stranded dna breaks https://academic.oup.com/nar/article-lookup/doi/10.1093/nar/gku172 doi:10.1093/nar/gku172 doi:10.1093/nar/gku172
spellingShingle mutation; dna; transposase; coordination; cytokinesis; dna transposons; double-stranded dna breaks
Claeys Bouuaert, Corentin
Walker, Neil
Liu, Danxu
Chalmers, Ronald
Crosstalk between transposase subunits during cleavage of the mariner transposon
title Crosstalk between transposase subunits during cleavage of the mariner transposon
title_full Crosstalk between transposase subunits during cleavage of the mariner transposon
title_fullStr Crosstalk between transposase subunits during cleavage of the mariner transposon
title_full_unstemmed Crosstalk between transposase subunits during cleavage of the mariner transposon
title_short Crosstalk between transposase subunits during cleavage of the mariner transposon
title_sort crosstalk between transposase subunits during cleavage of the mariner transposon
topic mutation; dna; transposase; coordination; cytokinesis; dna transposons; double-stranded dna breaks
url https://eprints.nottingham.ac.uk/41466/
https://eprints.nottingham.ac.uk/41466/
https://eprints.nottingham.ac.uk/41466/