title: Biochemical Characterization of the DASH-Type Cryptochrome CryD From Fusarium fujikuroi.
abstract: Proteins from the cryptochrome/photolyase family utilize UV-A, blue or even red light to achieve such diverse functions as repair of DNA lesions by photolyases and signaling by cryptochromes. DASH-type cryptochromes retained the ability to repair cyclobutane pyrimidine dimers (CPDs) in single-stranded DNA regions in vitro. However, most organisms possess conventional CPD photolyases responsible for repair of these lesions in vivo. Recent work showed that the DASH-type cryptochrome CryD plays a regulatory role in diverse light-dependent processes in Fusarium fujikuroi. Here, we report our in vitro studies on heterologously expressed FfCryD. The purified protein contains N(5) ,N(10) -methenyltetrahydrofolate and flavin adenine dinucleotide as cofactors. Photoreduction and DNA photorepair experiments confirmed that FfCryD is active in light-driven electron transfer processes. However, the protein showed comparable affinities for CPD-comprising and undamaged DNA probes. Surprisingly, after purification, full-length FfCryD as well as a truncated version containing only the PHR domain bound RNA which influenced their behavior in vitro. Moreover, binding of FfCryD to RNA indicates a putative role in RNA metabolism or in posttranscriptional control of gene expression.
authors.affiliation: Department of Genetics, Faculty of Biology, University of Seville, Seville, Spain.
authors.forename: Marta
authors.initials: M
authors.lastname: Castrillo
authors.affiliation: Department of Plant Physiology and Photobiology, Faculty of Biology, Philipps-University, Marburg, Germany.
authors.forename: Adrian
authors.initials: A
authors.lastname: Bernhardt
authors.affiliation: Department of Genetics, Faculty of Biology, University of Seville, Seville, Spain.
authors.forename: Javier
authors.initials: J
authors.lastname: Ávalos
authors.affiliation: Department of Plant Physiology and Photobiology, Faculty of Biology, Philipps-University, Marburg, Germany.
authors.forename: Alfred
authors.initials: A
authors.lastname: Batschauer
authors.affiliation: Department of Plant Physiology and Photobiology, Faculty of Biology, Philipps-University, Marburg, Germany.
authors.forename: Richard
authors.initials: R
authors.lastname: Pokorny
mesh_terms: D056931:Cryptochromes / Q000737:chemistry*; D004268:DNA-Binding Proteins / Q000737:chemistry; D005670:Fusarium / Q000737:chemistry*; D008027:Light; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D015398:Signal Transduction
chemical_list: D056931:Cryptochromes; D004268:DNA-Binding Proteins