Structural insights into the mechanism of double strand break formation by Hermes, a hAT family eukaryotic DNA transposase

Alison B. Hickman, Andrea Regier Voth, Hosam Ewis, Xianghong Li, Nancy L Craig, Fred Dyda

Research output: Contribution to journalArticle

Abstract

Some DNA transposons relocate from one genomic location to another using a mechanism that involves generating double-strand breaks at their transposon ends by forming hairpins on flanking DNA. The same double-strand break mode is employed by the V(D)J recombinase at signal-end/coding-end junctions during the generation of antibody diversity. How flanking hairpins are formed during DNA transposition has remained elusive. Here, we describe several co-crystal structures of the Hermes transposase bound to DNA that mimics the reaction step immediately prior to hairpin formation. Our results reveal a large DNA conformational change between the initial cleavage step and subsequent hairpin formation that changes which strand is acted upon by a single active site. We observed that two factors affect the conformational change: the complement of divalent metal ions bound by the catalytically essential DDE residues, and the identity of the -2 flanking base pair. Our data also provides a mechanistic link between the efficiency of hairpin formation (an A:T basepair is favored at the -2 position) and Hermes' strong target site preference. Furthermore, we have established that the histidine residue within a conserved C/DxxH motif present in many transposase families interacts directly with the scissile phosphate, suggesting a crucial role in catalysis.

Original languageEnglish (US)
Pages (from-to)10286-10301
Number of pages16
JournalNucleic Acids Research
Volume46
Issue number19
DOIs
StatePublished - Nov 2 2018

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Transposases
DNA
VDJ Recombinases
Antibody Diversity
Dichlorodiphenyl Dichloroethylene
DNA Transposable Elements
Catalysis
Histidine
Base Pairing
Catalytic Domain
Metals
Phosphates
Ions

ASJC Scopus subject areas

  • Genetics

Cite this

Structural insights into the mechanism of double strand break formation by Hermes, a hAT family eukaryotic DNA transposase. / Hickman, Alison B.; Voth, Andrea Regier; Ewis, Hosam; Li, Xianghong; Craig, Nancy L; Dyda, Fred.

In: Nucleic Acids Research, Vol. 46, No. 19, 02.11.2018, p. 10286-10301.

Research output: Contribution to journalArticle

Hickman, Alison B. ; Voth, Andrea Regier ; Ewis, Hosam ; Li, Xianghong ; Craig, Nancy L ; Dyda, Fred. / Structural insights into the mechanism of double strand break formation by Hermes, a hAT family eukaryotic DNA transposase. In: Nucleic Acids Research. 2018 ; Vol. 46, No. 19. pp. 10286-10301.
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