Affinities of Terminal Inverted Repeats to DNA Binding Domain of Transposase Affect the Transposition Activity of Bamboo Ppmar2 Mariner-Like Element

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Ramakrishnan , M , Zhou , M , Pan , C , Hanninen , H , Yrjala , K , Vinod , K K & Tang , D 2019 , ' Affinities of Terminal Inverted Repeats to DNA Binding Domain of Transposase Affect the Transposition Activity of Bamboo Ppmar2 Mariner-Like Element ' , International Journal of Molecular Sciences , vol. 20 , no. 15 , 3692 . https://doi.org/10.3390/ijms20153692

Title: Affinities of Terminal Inverted Repeats to DNA Binding Domain of Transposase Affect the Transposition Activity of Bamboo Ppmar2 Mariner-Like Element
Author: Ramakrishnan, Muthusamy; Zhou, Mingbing; Pan, Chunfang; Hanninen, Heikki; Yrjala, Kim; Vinod, Kunnummal Kurungara; Tang, Dingqin
Contributor: University of Helsinki, Department of Forest Sciences
Date: 2019-08
Language: eng
Number of pages: 16
Belongs to series: International Journal of Molecular Sciences
ISSN: 1661-6596
URI: http://hdl.handle.net/10138/305405
Abstract: Mariner-like elements (MLE) are a super-family of DNA transposons widespread in animal and plant genomes. Based on their transposition characteristics, such as random insertions and high-frequency heterogeneous transpositions, several MLEs have been developed to be used as tools in gene tagging and gene therapy. Two active MLEs, Ppmar1 and Ppmar2, have previously been identified in moso bamboo (Phyllostachys edulis). Both of these have a preferential insertion affinity to AT-rich region and their insertion sites are close to random in the host genome. In Ppmar2 element, we studied the affinities of terminal inverted repeats (TIRs) to DNA binding domain (DBD) and their influence on the transposition activity. We could identify two putative boxes in the TIRs which play a significant role in defining the TIR's affinities to the DBD. Seven mutated TIRs were constructed, differing in affinities based on similarities with those of other plant MLEs. Gel mobility shift assays showed that the TIR mutants with mutation sites G669A-C671A had significantly higher affinities than the mutants with mutation sites C657T-A660T. The high-affinity TIRs indicated that their transposition frequency was 1.5-2.0 times higher than that of the wild type TIRs in yeast transposition assays. The MLE mutants with low-affinity TIRs had relatively lower transposition frequency from that of wild types. We conclude that TIR affinity to DBD significantly affects the transposition activity of Ppmar2. The mutant MLEs highly active TIRs constructed in this study can be used as a tool for bamboo genetic studies.
Subject: Transposon
Mariner-like elements
Terminal inverted repeat
DNA binding domain
Transposase
Transposition activity
Phyllostachys edulis
Moso bamboo
SLEEPING-BEAUTY
GENOME
IDENTIFICATION
HYPERACTIVITY
4112 Forestry
1182 Biochemistry, cell and molecular biology
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