Factors responsible for target site selection in Tn10 transposition: a role for the DDE motif in target DNA capture

The EMBO Journal
M S Junop, D B Haniford

Abstract

Tn10, like several other transposons, exhibits a marked preference for integration into particular target sequences. Such sequences are referred to as integration hotspots and have been used to define a consensus target site in Tn10 transposition. We demonstrate that a Tn10 hotspot called HisG1, which was identified originally in vivo, also functions as an integration hotspot in vitro in a reaction where the HisG1 sequence is present on a short DNA oligomer. We use this in vitro system to define factors which are important for the capture of the HisG1 target site. We demonstrate that although divalent metal ions are not essential for HisG1 target capture, they greatly facilitate capture of a mutated HisG1 site. Analysis of catalytic transposase mutants further demonstrates that the DDE motif plays a critical role in 'divalent metal ion-dependent' target capture. Analysis of two other classes of transposase mutants, Exc+ Int- (which carry out transposon excision but not integration) and ATS (altered target specificity), demonstrates that while a particular ATS transposase binds HisG1 mutants better than wild-type transposase, Exc+ Int- mutants are defective in HisG1 capture, further defining the properties of these classes of mu...Continue Reading

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Citations

Dec 4, 2003·Journal of Molecular Biology·Ferenc OlaszJános Kiss
Jun 8, 2002·Journal of Molecular Biology·John S Allingham, David B Haniford
Jun 26, 2003·Journal of Molecular Biology·Patrick A Pribil, David B Haniford
Jun 2, 2001·The EMBO Journal·J S AllinghamD B Haniford
Aug 28, 2001·Nucleic Acids Research·L ZhangD J Finnegan
Aug 22, 2009·Nucleic Acids Research·Simon J WardleDavid B Haniford
Sep 17, 2005·Genes & Development·Simon J WardleDavid B Haniford
Jun 8, 2001·Molecular and Cellular Biology·M B NeiditchD B Roth
Jun 6, 2007·The Journal of Clinical Investigation·Marina Cavazzana-Calvo, Alain Fischer
Dec 19, 2008·Retrovirology·Olivier DelelisJean-François Mouscadet
Sep 2, 1998·Proceedings of the National Academy of Sciences of the United States of America·I Y GoryshinW S Reznikoff
Nov 10, 2006·Critical Reviews in Biochemistry and Molecular Biology·David B Haniford
Feb 17, 2005·Journal of Molecular Biology·Saima HumayunDavid B Haniford
Oct 4, 2006·Journal of Molecular Biology·Crystal R WhitfieldDavid B Haniford
Oct 20, 1998·Cell·A K KennedyD B Haniford
Nov 5, 1999·Annual Review of Microbiology·L HarenM Chandler
Feb 5, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Hatem ZayedZoltán Ivics
Feb 5, 2004·Molecular Therapy : the Journal of the American Society of Gene Therapy·Zsuzsanna Izsvák, Zoltán Ivics
Jun 20, 1998·The Journal of Biological Chemistry·S SantagataE Spanopoulou
Jun 22, 1999·Journal of Molecular Biology·J S AllinghamD B Haniford
Jun 25, 2015·Microbiology Spectrum·Zoltán Ivics, Zsuzsanna Izsvák

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