Functional characterization of the active Mutator-like transposable element, Muta1 from the mosquito Aedes aegyptiReport as inadecuate




Functional characterization of the active Mutator-like transposable element, Muta1 from the mosquito Aedes aegypti - Download this document for free, or read online. Document in PDF available to download.

Mobile DNA

, 8:1

First Online: 11 January 2017Received: 21 October 2016Accepted: 19 December 2016

Abstract

BackgroundMutator-like transposable elements MULEs are widespread with members in fungi, plants, and animals. Most of the research on the MULE superfamily has focused on plant MULEs where they were discovered and where some are extremely active and have significant impact on genome structure. The maize MuDR element has been widely used as a tool for both forward and reverse genetic studies because of its high transposition rate and preference for targeting genic regions. However, despite being widespread, only a few active MULEs have been identified, and only one, the rice Os3378, has demonstrated activity in a non-host organism.

ResultsHere we report the identification of potentially active MULEs in the mosquito Aedes aegypti. We demonstrate that one of these, Muta1, is capable of excision and reinsertion in a yeast transposition assay. Element reinsertion generated either 8 bp or 9 bp target site duplications TSDs with no apparent sequence preference. Mutagenesis analysis of donor site TSDs in the yeast assay indicates that their presence is important for precise excision and enhanced transposition. Site directed mutagenesis of the putative DDE catalytic motif and other conserved residues in the transposase protein abolished transposition activity.

ConclusionsCollectively, our data indicates that the Muta1 transposase of Ae. aegypti can efficiently catalyze both excision and reinsertion reactions in yeast. Mutagenesis analysis reveals that several conserved amino acids, including the DDE triad, play important roles in transposase function. In addition, donor site TSD also impacts the transposition of Muta1.

KeywordsTransposable elements Mutator-like elements MULE Aedes aegypti Yeast assay Target site duplication TSD Transposase AbbreviationsMULEMutator-like transposable elements

PCRPolymerase chain reaction

TETransposable element

TIRTerminal inverted repeat

TSDTarget site duplication

Electronic supplementary materialThe online version of this article doi:10.1186-s13100-016-0084-6 contains supplementary material, which is available to authorized users.

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Author: Kun Liu - Susan R. Wessler

Source: https://link.springer.com/







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