Temporal self-regulation of transposition through host-independent transposase rodlet formation

Transposons are highly abundant in eukaryotic genomes, but their mobilization must be finely tuned to maintain host organism fitness and allow for transposon propagation. Forty percent of the human genome is comprised of transposable element sequences, and the most abundant cut-and-paste transposons...

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Bibliographic Details
Published in:	Nucleic acids research Vol. 45; no. 1; pp. 353 - 366
Main Authors:	Woodard, Lauren E, Downes, Laura M, Lee, Yi-Chien, Kaja, Aparna, Terefe, Eyuel S, Wilson, Matthew H
Format:	Journal Article
Language:	English
Published:	England Oxford University Press 09-01-2017
Subjects:	Animals DNA Transposable Elements Female Gene Expression Regulation Genes, Reporter HEK293 Cells HeLa Cells Humans Insect Proteins - genetics Insect Proteins - metabolism Luciferases - genetics Luciferases - metabolism Male Mice Nucleic Acid Enzymes Optical Imaging Time-Lapse Imaging Transposases - genetics Transposases - metabolism Tribolium
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Summary:	Transposons are highly abundant in eukaryotic genomes, but their mobilization must be finely tuned to maintain host organism fitness and allow for transposon propagation. Forty percent of the human genome is comprised of transposable element sequences, and the most abundant cut-and-paste transposons are from the hAT superfamily. We found that the hAT transposase TcBuster from Tribolium castaneum formed filamentous structures, or rodlets, in human tissue culture cells, after gene transfer to adult mice, and ex vivo in cell-free conditions, indicating that host co-factors or cellular structures were not required for rodlet formation. Time-lapsed imaging of GFP-laced rodlets in human cells revealed that they formed quickly in a dynamic process involving fusion and fission. We delayed the availability of the transposon DNA and found that transposition declined after transposase concentrations became high enough for visible transposase rodlets to appear. In combination with earlier findings for maize Ac elements, these results give insight into transposase overproduction inhibition by demonstrating that the appearance of transposase protein structures and the end of active transposition are simultaneous, an effect with implications for genetic engineering and horizontal gene transfer.
Bibliography:	ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23
ISSN:	0305-1048 1362-4962
DOI:	10.1093/nar/gkw1115