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Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon
CRISPR-associated transposons (CASTs) are mobile genetic elements that co-opted CRISPR-Cas systems for RNA-guided transposition. Here we present the 2.4 Å cryo-EM structure of the Scytonema hofmannii (sh) TnsB transposase from Type V-K CAST, bound to the strand transfer DNA. The strand transfer comp...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9527255/ https://www.ncbi.nlm.nih.gov/pubmed/36184667 http://dx.doi.org/10.1038/s41467-022-33504-5 |
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author | Tenjo-Castaño, Francisco Sofos, Nicholas López-Méndez, Blanca Stutzke, Luisa S. Fuglsang, Anders Stella, Stefano Montoya, Guillermo |
author_facet | Tenjo-Castaño, Francisco Sofos, Nicholas López-Méndez, Blanca Stutzke, Luisa S. Fuglsang, Anders Stella, Stefano Montoya, Guillermo |
author_sort | Tenjo-Castaño, Francisco |
collection | PubMed |
description | CRISPR-associated transposons (CASTs) are mobile genetic elements that co-opted CRISPR-Cas systems for RNA-guided transposition. Here we present the 2.4 Å cryo-EM structure of the Scytonema hofmannii (sh) TnsB transposase from Type V-K CAST, bound to the strand transfer DNA. The strand transfer complex displays an intertwined pseudo-symmetrical architecture. Two protomers involved in strand transfer display a catalytically competent active site composed by DDE residues, while other two, which play a key structural role, show active sites where the catalytic residues are not properly positioned for phosphodiester hydrolysis. Transposon end recognition is accomplished by the NTD1/2 helical domains. A singular in trans association of NTD1 domains of the catalytically competent subunits with the inactive DDE domains reinforces the assembly. Collectively, the structural features suggest that catalysis is coupled to protein-DNA assembly to secure proper DNA integration. DNA binding residue mutants reveal that lack of specificity decreases activity, but it could increase transposition in some cases. Our structure sheds light on the strand transfer reaction of DDE transposases and offers new insights into CAST transposition. |
format | Online Article Text |
id | pubmed-9527255 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-95272552022-10-04 Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon Tenjo-Castaño, Francisco Sofos, Nicholas López-Méndez, Blanca Stutzke, Luisa S. Fuglsang, Anders Stella, Stefano Montoya, Guillermo Nat Commun Article CRISPR-associated transposons (CASTs) are mobile genetic elements that co-opted CRISPR-Cas systems for RNA-guided transposition. Here we present the 2.4 Å cryo-EM structure of the Scytonema hofmannii (sh) TnsB transposase from Type V-K CAST, bound to the strand transfer DNA. The strand transfer complex displays an intertwined pseudo-symmetrical architecture. Two protomers involved in strand transfer display a catalytically competent active site composed by DDE residues, while other two, which play a key structural role, show active sites where the catalytic residues are not properly positioned for phosphodiester hydrolysis. Transposon end recognition is accomplished by the NTD1/2 helical domains. A singular in trans association of NTD1 domains of the catalytically competent subunits with the inactive DDE domains reinforces the assembly. Collectively, the structural features suggest that catalysis is coupled to protein-DNA assembly to secure proper DNA integration. DNA binding residue mutants reveal that lack of specificity decreases activity, but it could increase transposition in some cases. Our structure sheds light on the strand transfer reaction of DDE transposases and offers new insights into CAST transposition. Nature Publishing Group UK 2022-10-02 /pmc/articles/PMC9527255/ /pubmed/36184667 http://dx.doi.org/10.1038/s41467-022-33504-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Tenjo-Castaño, Francisco Sofos, Nicholas López-Méndez, Blanca Stutzke, Luisa S. Fuglsang, Anders Stella, Stefano Montoya, Guillermo Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon |
title | Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon |
title_full | Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon |
title_fullStr | Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon |
title_full_unstemmed | Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon |
title_short | Structure of the TnsB transposase-DNA complex of type V-K CRISPR-associated transposon |
title_sort | structure of the tnsb transposase-dna complex of type v-k crispr-associated transposon |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9527255/ https://www.ncbi.nlm.nih.gov/pubmed/36184667 http://dx.doi.org/10.1038/s41467-022-33504-5 |
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