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© Melanie Blokesch, EPFL
Flagellated Vibrio cholerae
Publication : The EMBO journal

A target specificity switch in IS911 transposition: the role of the OrfA protein

Scientific Fields
Diseases
Organisms
Applications
Technique

Published in The EMBO journal - 01 Aug 2002

Loot C, Turlan C, Rousseau P, Ton-Hoang B, Chandler M

Link to Pubmed [PMID] – 12145217

EMBO J. 2002 Aug;21(15):4172-82

The role played by insertion sequence IS911 proteins, OrfA and OrfAB, in the choice of a target for insertion was studied. IS911 transposition occurs in several steps: synapsis of the two transposon ends (IRR and IRL); formation of a figure-of-eight intermediate where both ends are joined by a single-strand bridge; resolution into a circular form carrying an IRR-IRL junction; and insertion into a DNA target. In vivo, with OrfAB alone, an IS911-based transposon integrated with high probability next to an IS911 end located on the target plasmid. OrfA greatly reduced the proportion of these events. This was confirmed in vitro using a transposon with a preformed IRR-IRL junction to examine the final insertion step. Addition of OrfA resulted in a large increase in insertion frequency and greatly increased the proportion of non-targeted insertions. The intermolecular reaction leading to targeted insertion may resemble the intramolecular reaction involving figure-of-eight molecules, which leads to the formation of circles. OrfA could, therefore, be considered as a molecular switch modulating the site-specific recombination activity of OrfAB and facilitating dispersion of the insertion sequence (IS) to ‘non-homologous’ target sites.

http://www.ncbi.nlm.nih.gov/pubmed/12145217