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Structural Basis for the Inverted Repeat Preferences of mariner Transposases

The inverted repeat (IR) sequences delimiting the left and right ends of many naturally active mariner DNA transposons are non-identical and have different affinities for their transposase. We have compared the preferences of two active mariner transposases, Mos1 and Mboumar-9, for their imperfect t...

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Autores principales: Trubitsyna, Maryia, Grey, Heather, Houston, Douglas R., Finnegan, David J., Richardson, Julia M.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society for Biochemistry and Molecular Biology 2015
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4505599/
https://www.ncbi.nlm.nih.gov/pubmed/25869132
http://dx.doi.org/10.1074/jbc.M115.636704
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author Trubitsyna, Maryia
Grey, Heather
Houston, Douglas R.
Finnegan, David J.
Richardson, Julia M.
author_facet Trubitsyna, Maryia
Grey, Heather
Houston, Douglas R.
Finnegan, David J.
Richardson, Julia M.
author_sort Trubitsyna, Maryia
collection PubMed
description The inverted repeat (IR) sequences delimiting the left and right ends of many naturally active mariner DNA transposons are non-identical and have different affinities for their transposase. We have compared the preferences of two active mariner transposases, Mos1 and Mboumar-9, for their imperfect transposon IRs in each step of transposition: DNA binding, DNA cleavage, and DNA strand transfer. A 3.1 Å resolution crystal structure of the Mos1 paired-end complex containing the pre-cleaved left IR sequences reveals the molecular basis for the reduced affinity of the Mos1 transposase DNA-binding domain for the left IR as compared with the right IR. For both Mos1 and Mboumar-9, in vitro DNA transposition is most efficient when the preferred IR sequence is present at both transposon ends. We find that this is due to the higher efficiency of cleavage and strand transfer of the preferred transposon end. We show that the efficiency of Mboumar-9 transposition is improved almost 4-fold by changing the 3′ base of the preferred Mboumar-9 IR from guanine to adenine. This preference for adenine at the reactive 3′ end for both Mos1 and Mboumar-9 may be a general feature of mariner transposition.
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spelling pubmed-45055992015-07-17 Structural Basis for the Inverted Repeat Preferences of mariner Transposases Trubitsyna, Maryia Grey, Heather Houston, Douglas R. Finnegan, David J. Richardson, Julia M. J Biol Chem DNA and Chromosomes The inverted repeat (IR) sequences delimiting the left and right ends of many naturally active mariner DNA transposons are non-identical and have different affinities for their transposase. We have compared the preferences of two active mariner transposases, Mos1 and Mboumar-9, for their imperfect transposon IRs in each step of transposition: DNA binding, DNA cleavage, and DNA strand transfer. A 3.1 Å resolution crystal structure of the Mos1 paired-end complex containing the pre-cleaved left IR sequences reveals the molecular basis for the reduced affinity of the Mos1 transposase DNA-binding domain for the left IR as compared with the right IR. For both Mos1 and Mboumar-9, in vitro DNA transposition is most efficient when the preferred IR sequence is present at both transposon ends. We find that this is due to the higher efficiency of cleavage and strand transfer of the preferred transposon end. We show that the efficiency of Mboumar-9 transposition is improved almost 4-fold by changing the 3′ base of the preferred Mboumar-9 IR from guanine to adenine. This preference for adenine at the reactive 3′ end for both Mos1 and Mboumar-9 may be a general feature of mariner transposition. American Society for Biochemistry and Molecular Biology 2015-05-22 2015-04-13 /pmc/articles/PMC4505599/ /pubmed/25869132 http://dx.doi.org/10.1074/jbc.M115.636704 Text en © 2015 by The American Society for Biochemistry and Molecular Biology, Inc. Author's Choice—Final version free via Creative Commons CC-BY license (http://creativecommons.org/licenses/by/3.0) .
spellingShingle DNA and Chromosomes
Trubitsyna, Maryia
Grey, Heather
Houston, Douglas R.
Finnegan, David J.
Richardson, Julia M.
Structural Basis for the Inverted Repeat Preferences of mariner Transposases
title Structural Basis for the Inverted Repeat Preferences of mariner Transposases
title_full Structural Basis for the Inverted Repeat Preferences of mariner Transposases
title_fullStr Structural Basis for the Inverted Repeat Preferences of mariner Transposases
title_full_unstemmed Structural Basis for the Inverted Repeat Preferences of mariner Transposases
title_short Structural Basis for the Inverted Repeat Preferences of mariner Transposases
title_sort structural basis for the inverted repeat preferences of mariner transposases
topic DNA and Chromosomes
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4505599/
https://www.ncbi.nlm.nih.gov/pubmed/25869132
http://dx.doi.org/10.1074/jbc.M115.636704
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