Abstract
Most DNA transposons move from one genomic location to another by a cut-and-paste mechanism and are useful tools for genomic manipulations. Short inverted repeat (IR) DNA sequences marking each end of the transposon are recognized by a DNA transposase (encoded by the transposon itself). This enzyme cleaves the transposon ends and integrates them at a new genomic location. We report here a comparison of the biophysical and biochemical properties of two closely related and active mariner/Tc1 family DNA transposases: Mboumar-9 and Mos1. We compared the in vitro cleavage activities of the enzymes on their own IR sequences, as well as cross-recognition of their inverted repeat sequences. We found that, like Mos1, untagged recombinant Mboumar-9 transposase is a dimer and forms a stable complex with inverted repeat DNA in the presence of Mg2+ ions. Mboumar-9 transposase cleaves its inverted repeat DNA in the manner observed for Mos1 transposase. There was minimal cross-recognition of IR sequences between Mos1 and Mboumar-9 transposases, despite these enzymes having 68% identical amino acid sequences. Transposases sharing common biophysical and biochemical properties, but retaining recognition specificity toward their own IR, are a promising platform for the design of chimeric transposases with predicted and improved sequence recognition.
Highlights
To date, five naturally active eukaryotic mariner/Tc1 family elements have been described: Mos[1] from Drosophila mauritiana,[4] Famar[1] from the earwig Forf icula auriculata,[5] Mboumar-9 from the ant Messor bouvieri,[6] Minos from the fly Drosophila hydei,[7] and Passport from the plaice Pleuronectes platessa.[8]
Sequence comparisons reveal that Mos[1] and Mboumar-9 transposases are the most closely related active mariner transposases described to date, and they share 68% identical amino acid sequences[6,28] (Figure 1a)
Like Mos[1], the enzyme forms a dimer in solution, and this was recently established as a prerequisite for autoregulation in mariner transposition.[30]
Summary
Five naturally active eukaryotic mariner/Tc1 family elements have been described: Mos[1] from Drosophila mauritiana,[4] Famar[1] from the earwig Forf icula auriculata,[5] Mboumar-9 from the ant Messor bouvieri,[6] Minos from the fly Drosophila hydei,[7] and Passport from the plaice Pleuronectes platessa.[8]. Our results suggest that these closely related enzymes could be used to design chimeric proteins with predictable DNA recognition properties and enhanced efficiencies as genomic manipulation tools
Sign-in/Register to view highlights & summary 
Published Version (
Free)
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
