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

Nucleic Acids Research
Alison Burgess HickmanFred Dyda

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 ma...Continue Reading

References

Jul 15, 1993·Proceedings of the National Academy of Sciences of the United States of America·T A Steitz, J A Steitz
Oct 20, 1998·Cell·A K KennedyD B Haniford
Feb 17, 2001·The EMBO Journal·P N KuduvalliN L Craig
Jun 26, 2003·Journal of Molecular Biology·Patrick A Pribil, David B Haniford
Dec 20, 2003·Nature Reviews. Molecular Cell Biology·M Joan Curcio, Keith M Derbyshire
May 19, 2005·PLoS Biology·Vladimir V Kapitonov, Jerzy Jurka
Jul 26, 2005·Nature Structural & Molecular Biology·Alison B HickmanFred Dyda
Mar 21, 2008·The EMBO Journal·Rupak MitraNancy L Craig
Aug 6, 2008·Annual Review of Genetics·William S Reznikoff
Jan 24, 2009·EMBO Reports·Marcin Nowotny
Jan 14, 2010·Critical Reviews in Biochemistry and Molecular Biology·Alison Burgess HickmanFred Dyda
Mar 25, 2010·Heredity·D J RankinS P Brown
Dec 7, 2010·Proceedings of the National Academy of Sciences of the United States of America·Sunil GangadharanNancy L Craig
Mar 4, 2011·Genetics·Peter ArensburgerPeter W Atkinson
Apr 27, 2011·Proceedings of the National Academy of Sciences of the United States of America·Yao-Wu Yuan, Susan R Wessler
May 31, 2011·The Journal of Biological Chemistry·Richard A KatzAnna Marie Skalka
May 15, 2012·The EMBO Journal·Stephen HarePeter Cherepanov
Jul 10, 2012·Nature Structural & Molecular Biology·Cary G HenckenNancy L Craig
Oct 24, 2012·Proceedings of the National Academy of Sciences of the United States of America·Xianghong LiNancy L Craig
Nov 9, 2012·Nature·Sherwin P MontañoPhoebe A Rice
Nov 26, 2013·The Journal of Biological Chemistry·Aude PfliegerSylvaine Renault
Feb 7, 2014·FEMS Microbiology Reviews·Patricia SiguierMick Chandler
Mar 14, 2014·Nucleic Acids Research·Corentin Claeys BouuaertRonald Chalmers
Oct 15, 2014·Proceedings of the National Academy of Sciences of the United States of America·Naghmeh RajaeiStefan U Aström
Jan 16, 2015·Accounts of Chemical Research·Giulia PalermoMarco De Vivo
Feb 11, 2015·Nucleic Acids Research·Jacqueline DornanJulia M Richardson
Feb 25, 2015·Nature·Min-Sung KimMartin Gellert
May 1, 2015·Biology Direct·Vladimir V Kapitonov, Eugene V Koonin

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Methods Mentioned

BETA
X-ray
size exclusion chromatography
PCR

Software Mentioned

AMoRe
XDS
Phenix

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