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Kinetic Analysis of the Interaction of Mos1 Transposase with its Inverted Terminal Repeats Reveals New Insight into the Protein–DNA Complex Assembly
Author(s) -
Esnault Charles,
Jaillet Jérôme,
Delorme Nicolas,
Bouchet Nicolas,
Renault Sylvaine,
DouziechEyrolles Laurence,
Pilard JeanFrançois,
AugéGouillou Corinne
Publication year - 2015
Publication title -
chembiochem
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.05
H-Index - 126
eISSN - 1439-7633
pISSN - 1439-4227
DOI - 10.1002/cbic.201402466
Subject(s) - transposase , transposition (logic) , dna , inverted repeat , transposable element , chemistry , biology , genetics , gene , computer science , genome , artificial intelligence
Transposases are specific DNA‐binding proteins that promote the mobility of discrete DNA segments. We used a combination of physicochemical approaches to describe the association of MOS1 (an eukaryotic transposase) with its specific target DNA, an event corresponding to the first steps of the transposition cycle. Because the kinetic constants of the reaction are still unknown, we aimed to determine them by using quartz crystal microbalance on two sources of recombinant MOS1: one produced in insect cells and the other produced in bacteria. The prokaryotic‐expressed MOS1 showed no cooperativity and displayed a K d of about 300 n M . In contrast, the eukaryotic‐expressed MOS1 generated a cooperative system, with a lower K d (∼2 n m) . The origins of these differences were investigated by IR spectroscopy and AFM imaging. Both support the conclusion that prokaryotic‐ and eukaryotic‐expressed MOS1 are not similarly folded, thereby resulting in differences in the early steps of transposition.