title: Sleeping Beauty transposase structure allows rational design of hyperactive variants for genetic engineering.
abstract: Sleeping Beauty (SB) is a prominent Tc1/mariner superfamily DNA transposon that provides a popular genome engineering tool in a broad range of organisms. It is mobilized by a transposase enzyme that catalyses DNA cleavage and integration at short specific sequences at the transposon ends. To facilitate SB's applications, here we determine the crystal structure of the transposase catalytic domain and use it to model the SB transposase/transposon end/target DNA complex. Together with biochemical and cell-based transposition assays, our structure reveals mechanistic insights into SB transposition and rationalizes previous hyperactive transposase mutations. Moreover, our data enables us to design two additional hyperactive transposase variants. Our work provides a useful resource and proof-of-concept for structure-based engineering of tailored SB transposases.
authors.affiliation: European Molecular Biology Laboratory, Structural and Computational Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany.
authors.forename: Franka
authors.initials: F
authors.lastname: Voigt
authors.affiliation: Paul Ehrlich Institute, Division of Medical Biotechnology, Paul Ehrlich Strasse 51-59, Langen 63225, Germany.
authors.forename: Lisa
authors.initials: L
authors.lastname: Wiedemann
authors.affiliation: European Molecular Biology Laboratory, Structural and Computational Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany.
authors.forename: Cecilia
authors.initials: C
authors.lastname: Zuliani
authors.affiliation: European Molecular Biology Laboratory, Structural and Computational Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany.
authors.forename: Irma
authors.initials: I
authors.lastname: Querques
authors.affiliation: Paul Ehrlich Institute, Division of Medical Biotechnology, Paul Ehrlich Strasse 51-59, Langen 63225, Germany.
authors.forename: Attila
authors.initials: A
authors.lastname: Sebe
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert Rössle Strasse 10, Berlin 13092, Germany.
authors.forename: Lajos
authors.initials: L
authors.lastname: Mátés
authors.affiliation: Max Delbrück Center for Molecular Medicine, Robert Rössle Strasse 10, Berlin 13092, Germany.
authors.forename: Zsuzsanna
authors.initials: Z
authors.lastname: Izsvák
authors.affiliation: Paul Ehrlich Institute, Division of Medical Biotechnology, Paul Ehrlich Strasse 51-59, Langen 63225, Germany.
authors.forename: Zoltán
authors.initials: Z
authors.lastname: Ivics
authors.affiliation: European Molecular Biology Laboratory, Structural and Computational Biology Unit, Meyerhofstrasse 1, Heidelberg 69117, Germany.
authors.forename: Orsolya
authors.initials: O
authors.lastname: Barabas
mesh_terms: D020134:Catalytic Domain; D018360:Crystallography, X-Ray; D004251:DNA Transposable Elements; D005818:Genetic Engineering; D008958:Models, Molecular; D016296:Mutagenesis; D009154:Mutation / Q000235:genetics*; D019895:Transposases / Q000737:chemistry* / Q000378:metabolism*
chemical_list: D004251:DNA Transposable Elements; D019895:Transposases