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Characterization of a Tn5 pre-cleavage synaptic complex.

Abstract：

:Protein catalyzed DNA rearrangements typically require assembly of complex nucleoprotein structures. In transposition and integration reactions, these structures, termed synaptic complexes, are mandatory for catalysis. We characterize the Tn5 pre-cleavage synaptic complex, the simplest transposition complex described to date. We identified this complex by gel retardation assay using short, linear fragments and have shown that it contains a dimer of transposase, two DNA molecules, and is competent for DNA cleavage in the presence of Mg(2+). We also used hydroxyl radical footprinting and interference techniques to delineate the protein-DNA contacts made in the Tn5 synaptic and monomer complexes. All positions (except position 1) of the end sequence are contacted by transposase in the synaptic complex. We have determined that positions 2-5 of the end sequence are specifically required for synaptic complex formation as they are not required for monomer complex formation. In addition, in the synaptic complex, there is a strong, local distortion centered around position 1 which likely facilitates cleavage.

journal_name

J Mol Biol

journal_title

Journal of molecular biology

authors

Bhasin A,Goryshin IY,Steiniger-White M,York D,Reznikoff WS

doi

10.1006/jmbi.2000.4048

keywords:

subject

Has Abstract

pub_date

2000-09-08 00:00:00

pages

49-63

issue

1

eissn

0022-2836

issn

1089-8638

pii

S0022-2836(00)94048-8

journal_volume

302

pub_type

杂志文章

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