Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends
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...
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Nature Portfolio
2025-01-01
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| Series: | Nature Communications |
| Online Access: | https://doi.org/10.1038/s41467-024-55784-9 |
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| author | 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 |
| author_facet | 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 |
| author_sort | Alison B. Hickman |
| collection | DOAJ |
| description | 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. |
| format | Article |
| id | doaj-art-84fe65d9e7f44e3e8cd6e66dc0020bf5 |
| institution | Kabale University |
| issn | 2041-1723 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | Nature Portfolio |
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| series | Nature Communications |
| spelling | doaj-art-84fe65d9e7f44e3e8cd6e66dc0020bf52025-01-12T12:31:51ZengNature PortfolioNature Communications2041-17232025-01-0116111810.1038/s41467-024-55784-9Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon endsAlison B. Hickman0Laurie Lannes1Christopher M. Furman2Christina Hong3Lidiya Franklin4Rodolfo Ghirlando5Arpita Ghosh6Wentian Luo7Parthena Konstantinidou8Hernán A. Lorenzi9Anne Grove10Astrid D. Haase11Matthew H. Wilson12Fred Dyda13Laboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthLaboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthLaboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthLaboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthLaboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthLaboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthDepartment of Biological Sciences, Louisiana State UniversityDepartment of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical CenterLaboratory of Cellular and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthLaboratory of Biochemistry and Genetics, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthDepartment of Biological Sciences, Louisiana State UniversityLaboratory of Cellular and Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthDepartment of Medicine, Division of Nephrology and Hypertension, Vanderbilt University Medical CenterLaboratory of Molecular Biology, National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of HealthAbstract 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.https://doi.org/10.1038/s41467-024-55784-9 |
| spellingShingle | 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 Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends Nature Communications |
| title | Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends |
| title_full | Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends |
| title_fullStr | Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends |
| title_full_unstemmed | Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends |
| title_short | Activity of the mammalian DNA transposon piggyBat from Myotis lucifugus is restricted by its own transposon ends |
| title_sort | activity of the mammalian dna transposon piggybat from myotis lucifugus is restricted by its own transposon ends |
| url | https://doi.org/10.1038/s41467-024-55784-9 |
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