title: Programmable DNA-binding proteins from Burkholderia provide a fresh perspective on the TALE-like repeat domain.
abstract: The tandem repeats of transcription activator like effectors (TALEs) mediate sequence-specific DNA binding using a simple code. Naturally, TALEs are injected by Xanthomonas bacteria into plant cells to manipulate the host transcriptome. In the laboratory TALE DNA binding domains are reprogrammed and used to target a fused functional domain to a genomic locus of choice. Research into the natural diversity of TALE-like proteins may provide resources for the further improvement of current TALE technology. Here we describe TALE-like proteins from the endosymbiotic bacterium Burkholderia rhizoxinica, termed Bat proteins. Bat repeat domains mediate sequence-specific DNA binding with the same code as TALEs, despite less than 40% sequence identity. We show that Bat proteins can be adapted for use as transcription factors and nucleases and that sequence preferences can be reprogrammed. Unlike TALEs, the core repeats of each Bat protein are highly polymorphic. This feature allowed us to explore alternative strategies for the design of custom Bat repeat arrays, providing novel insights into the functional relevance of non-RVD residues. The Bat proteins offer fertile grounds for research into the creation of improved programmable DNA-binding proteins and comparative insights into TALE-like evolution.
authors.affiliation: Genetics, Department of Biology I, Ludwig-Maximilians-University Munich, Martinsried, Bavaria, 82152, Germany.
authors.forename: Orlando
authors.initials: O
authors.lastname: de Lange
authors.affiliation: Genetics, Department of Biology I, Ludwig-Maximilians-University Munich, Martinsried, Bavaria, 82152, Germany.
authors.forename: Christina
authors.initials: C
authors.lastname: Wolf
authors.affiliation: Genetics, Department of Biology I, Ludwig-Maximilians-University Munich, Martinsried, Bavaria, 82152, Germany.
authors.forename: Jörn
authors.initials: J
authors.lastname: Dietze
authors.affiliation: Genetics, Department of Biology I, Ludwig-Maximilians-University Munich, Martinsried, Bavaria, 82152, Germany.
authors.forename: Janett
authors.initials: J
authors.lastname: Elsaesser
authors.affiliation: Genetics, Department of Biology I, Ludwig-Maximilians-University Munich, Martinsried, Bavaria, 82152, Germany.
authors.forename: Robert
authors.initials: R
authors.lastname: Morbitzer
authors.affiliation: Genetics, Department of Biology I, Ludwig-Maximilians-University Munich, Martinsried, Bavaria, 82152, Germany thomas.lahaye@zmbp.uni-tuebingen.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Lahaye
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D019117:Burkholderia / Q000235:genetics*; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D053837:DNA Cleavage; D004268:DNA-Binding Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000737:chemistry / Q000378:metabolism; D016680:Genome, Bacterial; D057809:HEK293 Cells; D006801:Humans; D019913:Nuclear Localization Signals; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D020449:Repetitive Sequences, Amino Acid; D015534:Trans-Activators / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D015533:Transcriptional Activation
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; D019913:Nuclear Localization Signals; D015534:Trans-Activators; D004247:DNA; C035042:endodeoxyribonuclease FokI; D015252:Deoxyribonucleases, Type II Site-Specific