Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends

Alison B Hickman; Laurie Lannes; Christopher M Furman; Christina Hong; Lidiya Franklin; Rodolfo Ghirlando; Arpita Ghosh; Wentian Luo; Parthena Konstantinidou; Hernán A Lorenzi; Anne Grove; Astrid D Haase; Matthew H Wilson; Fred Dyda

doi:10.1038/s41467-024-55784-9

Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends

Nat Commun. 2025 Jan 7;16(1):458. doi: 10.1038/s41467-024-55784-9.

Authors

Alison B Hickman¹, Laurie Lannes^#^{1

2}, Christopher M Furman^#^{1

3}, Christina Hong¹, Lidiya Franklin¹, Rodolfo Ghirlando¹, Arpita Ghosh⁴, Wentian Luo⁵, Parthena Konstantinidou⁶, Hernán A Lorenzi⁷, Anne Grove⁴, Astrid D Haase⁶, Matthew H Wilson^{5

8

9

10}, Fred Dyda¹¹

Affiliations

¹ Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
² Structural Motility, UMR 144 CNRS/Curie Institute, PSL Research University, Paris, cedex 05, France.
³ International Flavors and Fragrances, Wilmington, DE, USA.
⁴ Department of Biological Sciences, Louisiana State University, Baton Rouge, LA, USA.
⁵ Department of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical Center, Nashville, TN, USA.
⁶ Laboratory of Cellular and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
⁷ Laboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA.
⁸ Department of Veterans Affairs, Nashville, TN, USA.
⁹ Department of Pharmacology, Vanderbilt University, Nashville, TN, USA.
¹⁰ Department of Cell and Developmental Biology, Vanderbilt University, Nashville, TN, USA.
¹¹ Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, Bethesda, MD, USA. [email protected].

^# Contributed equally.

PMID: 39774116
DOI: 10.1038/s41467-024-55784-9

Abstract

Members of the piggyBac superfamily of DNA transposons are widely distributed in host genomes ranging from insects to mammals. The human genome has retained five piggyBac-derived genes as domesticated elements although they are no longer mobile. Here, we have investigated the transposition properties of piggyBat from Myotis lucifugus, the only known active mammalian DNA transposon, and show that its low activity in human cells is due to subterminal inhibitory DNA sequences. Activity can be dramatically improved by their removal, suggesting the existence of a mechanism for the suppression of transposon activity. The cryo-electron microscopy structure of the piggyBat transposase pre-synaptic complex showed an unexpected mode of DNA binding and recognition using C-terminal domains that are topologically different from those of the piggyBac transposase. Here we show that structure-based rational re-engineering of the transposase through the removal of putative phosphorylation sites and a changed domain organization - in combination with truncated transposon ends - results in a transposition system that is at least 100-fold more active than wild-type piggyBat.

MeSH terms

Animals
Cryoelectron Microscopy
DNA / genetics
DNA / metabolism
DNA Transposable Elements* / genetics
Humans
Transposases* / genetics
Transposases* / metabolism

Substances

DNA Transposable Elements
Transposases
DNA

Abstract

MeSH terms

Substances

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