Journal Title
Title of Journal: Genetica
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Publisher
Springer Netherlands
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Authors: Stéphanie Germon Nicolas Bouchet Sophie Casteret Guillaume Carpentier Jérémy Adet Yves Bigot Corinne AugéGouillou
Publish Date: 2009/06/17
Volume: 137, Issue: 3, Pages: 265-
Abstract
Mariner transposons are probably the most widespread transposable element family in animal genomes To date they are believed not to require speciesspecific host factors for transposition Despite this Mos1 one of the moststudied mariner elements with Himar1 has been shown to be active in insects but inactive in mammalian genomes To circumvent this problem one strategy consists of both enhancing the activity of the Mos1 transposase MOS1 and making it insensitive to activityaltering posttranslational modifications Here we report rational mutagenesis studies performed to obtain hyperactive and nonphosphorylable MOS1 variants Transposition assays in bacteria have made it possible to isolate numerous hyperactive MOS1 variants The best mutant combinations named FETY and FET are 60 and 800fold more active than the wildtype MOS1 version respectively However there are serious difficulties in using them notably because they display severe cytotoxicity On the other hand three positions lying within the HTH motif T88 S99 and S104 were found to be sensitive to phosphorylation Our efforts to obtain active nonphosphorylable mutants at S99 and S104 positions were unsuccessful as these residues like the colinear amino acids in their close vicinity are critical for MOS1 activity Even if host factors are not essential for transposition our data demonstrate that the host machinery is essential in regulating MOS1 activityThis work was funded by the University of Tours the Centre National de la Recherche Scientifique the Ministère de l’Education Nationale de la Recherche et de la Technologie and GDR CNRS no 2157 This work was also funded by grants from the European Commission Project SyntheGeneDelivery grant no 018716 to YB the Association Française contre les Myopathies grant AFM no 11468 to CAG and the Agence Nationale de la Recherche Project Magic Transpo grant ANR06EMPB01401 to CAG
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