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Common physical properties of DNA affecting target site selection of sleeping beauty and other Tc1/mariner transposable elements.

Abstract：

:Sleeping Beauty (SB) is the most active Tc1/mariner-type transposable element in vertebrates, and is therefore a valuable vector for transposon mutagenesis in vertebrate models and for human gene therapy. We have analyzed factors affecting target site selection of SB in mammalian cells, by generating transposition events from extrachromosomal plasmids to chromosomes. In contrast to the local hopping observed when transposition is induced from a chromosomal context, mapping of 138 unique SB insertions on human chromosomes showed a fairly random genomic distribution, and a 35% occurrence of transposition into genes. Inspection of the DNA flanking the sites of element integration revealed significant differences from random DNA in both primary sequence and physical properties. The consensus sequence of SB target sites was found to be a palindromic AT-repeat, ATATATAT, in which the central TA is the canonical target site. We found however, that target site selection is determined primarily on the level of DNA structure, and not by specific base-pair interactions. Computational analyses revealed that insertion sites tend to have a bendable structure and a palindromic pattern of potential hydrogen-bonding sites in the major groove of the DNA. These features appear conserved in the Tc1/mariner family of transposons and in other, distantly related elements that share a common catalytic domain of the transposase, and integrate fairly randomly. No similar target site preference was found for non-randomly integrating elements. Our results suggest common factors influencing target site selection of a wide range of transposable elements.

journal_name

J Mol Biol

journal_title

Journal of molecular biology

authors

Vigdal TJ,Kaufman CD,Izsvák Z,Voytas DF,Ivics Z

doi

10.1016/s0022-2836(02)00991-9

keywords:

subject

Has Abstract

pub_date

2002-10-25 00:00:00

pages

441-52

issue

3

eissn

0022-2836

issn

1089-8638

pii

S0022283602009919

journal_volume

323

pub_type

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