Structural role of the flanking DNA in mariner transposon excision

Jacqueline Dornan, Heather Grey, Julia M Richardson*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

Abstract

During cut-and-paste mariner/Tc1 transposition, transposon DNA is cut precisely at its junction with flanking DNA, ensuring the transposon is neither shortened nor lengthened with each transposition event. Each transposon end is flanked by a TpA dinucleotide: the signature target site duplication of mariner/Tc1 transposition. To establish the role of this sequence in accurate DNA cleavage, we have determined the crystal structure of a pre-second strand cleavage mariner Mos1 transpososome. The structure reveals the route of an intact DNA strand through the transposase active site before second strand cleavage. The crossed architecture of this pre-second strand cleavage paired-end complex supports our proposal that second strand cleavage occurs in trans. The conserved mariner transposase WVPHEL and YSPDL motifs position the strand for accurate DNA cleavage. Base-specific recognition of the flanking DNA by conserved amino acids is revealed, defining a new role for the WVPHEL motif in mariner transposition and providing a molecular explanation for in vitro mutagenesis data. Comparison of the pre-TS cleavage and post-cleavage Mos1 transpososomes with structures of Prototype Foamy Virus intasomes suggests a binding mode for target DNA prior to Mos1 transposon integration.

Original languageEnglish
Pages (from-to)2424-2432
Number of pages9
JournalNucleic Acids Research
Volume43
Issue number4
Early online date8 Feb 2015
DOIs
Publication statusPublished - 27 Feb 2015

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  • DNA transactions

    Julia Richardson (Invited speaker)

    7 Sep 201412 Sep 2014

    Activity: Participating in or organising an event typesParticipation in conference

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