Structures of ISCth4 transpososomes reveal the role of asymmetry in copy-out/paste-in DNA transposition.
antibiotic resistance
crystallography
mechanism
promoter
transposon
Journal
The EMBO journal
ISSN: 1460-2075
Titre abrégé: EMBO J
Pays: England
ID NLM: 8208664
Informations de publication
Date de publication:
04 01 2021
04 01 2021
Historique:
received:
19
05
2020
revised:
07
08
2020
accepted:
10
09
2020
pubmed:
3
10
2020
medline:
21
4
2021
entrez:
2
10
2020
Statut:
ppublish
Résumé
Copy-out/paste-in transposition is a major bacterial DNA mobility pathway. It contributes significantly to the emergence of antibiotic resistance, often by upregulating expression of downstream genes upon integration. Unlike other transposition pathways, it requires both asymmetric and symmetric strand transfer steps. Here, we report the first structural study of a copy-out/paste-in transposase and demonstrate its ability to catalyze all pathway steps in vitro. X-ray structures of ISCth4 transposase, a member of the IS256 family of insertion sequences, bound to DNA substrates corresponding to three sequential steps in the reaction reveal an unusual asymmetric dimeric transpososome. During transposition, an array of N-terminal domains binds a single transposon end while the catalytic domain moves to accommodate the varying substrates. These conformational changes control the path of DNA flanking the transposon end and the generation of DNA-binding sites. Our results explain the asymmetric outcome of the initial strand transfer and show how DNA binding is modulated by the asymmetric transposase to allow the capture of a second transposon end and to integrate a circular intermediate.
Identifiants
pubmed: 33006208
doi: 10.15252/embj.2020105666
pmc: PMC7780238
doi:
Substances chimiques
DNA Transposable Elements
0
DNA, Bacterial
0
DNA-Binding Proteins
0
Transposases
EC 2.7.7.-
Banques de données
PDB
['6XG8', '6XGW', '6XGX']
GENBANK
['CP000568.1']
Types de publication
Journal Article
Research Support, N.I.H., Extramural
Research Support, N.I.H., Intramural
Research Support, U.S. Gov't, Non-P.H.S.
Langues
eng
Sous-ensembles de citation
IM
Pagination
e105666Subventions
Organisme : Intramural NIH HHS
ID : Z01 DK036154
Pays : United States
Organisme : NCRR NIH HHS
ID : S10 RR028976
Pays : United States
Organisme : NCRR NIH HHS
ID : S10 RR025528
Pays : United States
Informations de copyright
Published 2020. This article is a U.S. Government work and is in the public domain in the USA.
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