Nucleic Acids Research, Vol 24, Issue 19 3790-3796, Copyright © 1996 by Oxford University Press
D York and WS Reznikoff
The binding of transposase (Tnp) to the specific Tn5 end sequences is the
first dedicated reaction during transposition. In this study, comparative
DNA-binding analyses were performed using purified full- length Tnp and a
C-terminal deletion variant (delta369) that lacks the putative dimerization
domain. The shape of the binding curve of full- length Tnp is sigmoidal in
contrast to the hyperbolic-shaped binding curve of delta369. This
observation is consistent with previous observations as well as a rate of
binding study presented here, which suggest that the full-length Tnp-end
interaction, unlike that of the truncated protein, is a complex
time-dependent reaction possibly involving a subunit exchange. Circular
permutation assay results indicate that both proteins are capable of
distorting the Tn5end sequences upon binding. Molecular weight
determinations based on the migratory patterns of complexed DNA in
polyacrylamide gels has shown that delta369 specifically binds the Tn5 end
sequences as a monomer while full-length Tnp in complex represents a
heterodimer.
ARTICLES
Purification and biochemical analyses of a monomeric form of Tn5 transposase
Department of Biochemistry, University of Wisconsin-Madison, 53706, USA.
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