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TitleStructural basis of seamless excision and specific targeting by piggyBac transposase.
Journal, issue, pagesNat Commun, Vol. 11, Issue 1, Page 3446, Year 2020
Publish dateJul 10, 2020
AuthorsQiujia Chen / Wentian Luo / Ruth Ann Veach / Alison B Hickman / Matthew H Wilson / Fred Dyda /
PubMed AbstractThe piggyBac DNA transposon is used widely in genome engineering applications. Unlike other transposons, its excision site can be precisely repaired without leaving footprints and it integrates ...The piggyBac DNA transposon is used widely in genome engineering applications. Unlike other transposons, its excision site can be precisely repaired without leaving footprints and it integrates specifically at TTAA tetranucleotides. We present cryo-EM structures of piggyBac transpososomes: a synaptic complex with hairpin DNA intermediates and a strand transfer complex capturing the integration step. The results show that the excised TTAA hairpin intermediate and the TTAA target adopt essentially identical conformations, providing a mechanistic link connecting the two unique properties of piggyBac. The transposase forms an asymmetric dimer in which the two central domains synapse the ends while two C-terminal domains form a separate dimer that contacts only one transposon end. In the strand transfer structure, target DNA is severely bent and the TTAA target is unpaired. In-cell data suggest that asymmetry promotes synaptic complex formation, and modifying ends with additional transposase binding sites stimulates activity.
External linksNat Commun / PubMed:32651359 / PubMed Central
MethodsEM (single particle)
Resolution3.47 - 3.66 Å
Structure data

22072

6x67

EMDB-22072, PDB-6x67:
Cryo-EM structure of piggyBac transposase strand transfer complex (STC)
Method: EM (single particle) / Resolution: 3.47 Å

22073

6x68

EMDB-22073, PDB-6x68:
Cryo-EM structure of piggyBac transposase synaptic complex with hairpin DNA (SNHP)
Method: EM (single particle) / Resolution: 3.66 Å

Chemicals

ChemComp-ZN:
Unknown entry

ChemComp-CA:
Unknown entry

Source
  • trichoplusia ni (cabbage looper)
KeywordsDNA BINDING PROTEIN/DNA / piggyBac / strand transfer complex / transpososome / target DNA / DNA BINDING PROTEIN-DNA complex / synaptic complex / hairpin DNA / transposase

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