title: The Myb domain of LUX ARRHYTHMO in complex with DNA: expression, purification and crystallization.
abstract: LUX ARRHYTHMO (LUX) is a Myb-domain transcription factor that plays an important role in regulating the circadian clock. Lux mutations cause severe clock defects and arrhythmia in constant light and dark. In order to examine the molecular mechanisms underlying the function of LUX, the DNA-binding Myb domain was cloned, expressed and purified. The DNA-binding activity of the Myb domain was confirmed using electrophoretic mobility shift assays (EMSAs), demonstrating that the LUX Myb domain is able to bind to DNA with nanomolar affinity. In order to investigate the specificity determinants of protein-DNA interactions, the protein was co-crystallized with a 10-mer cognate DNA. Initial crystallization results for the selenomethionine-derivatized protein and data-set collection statistics are reported. Data collection was performed using the MeshAndCollect workflow available at the ESRF.
authors.affiliation: Laboratoire de Physiologie Cellulaire et Vegetale, Biosciences and Biotechnology Institute of Grenoble, UMR5168, CNRS/CEA/INRA/UGA, 17 Rue des Martyrs, 38054 Grenoble, France.
authors.forename: Catarina S
authors.initials: CS
authors.lastname: Silva
authors.affiliation: Structural Biology Group, European Synchrotron Radiation Facility, 71 Avenue des Martyrs, 38054 Grenoble, France.
authors.forename: Xuelei
authors.initials: X
authors.lastname: Lai
authors.affiliation: European Molecular Biology Laboratory, 71 Avenue des Martyrs, 38054 Grenoble, France.
authors.forename: Max
authors.initials: M
authors.lastname: Nanao
authors.affiliation: Laboratoire de Physiologie Cellulaire et Vegetale, Biosciences and Biotechnology Institute of Grenoble, UMR5168, CNRS/CEA/INRA/UGA, 17 Rue des Martyrs, 38054 Grenoble, France.
authors.forename: Chloe
authors.initials: C
authors.lastname: Zubieta
mesh_terms: D017360:Arabidopsis / Q000737:chemistry* / Q000235:genetics; D029681:Arabidopsis Proteins / Q000737:chemistry* / Q000302:isolation & purification; D003460:Crystallization; D018360:Crystallography, X-Ray; D004247:DNA / Q000737:chemistry* / Q000302:isolation & purification; D024202:Electrophoretic Mobility Shift Assay; D017343:Genes, Plant; D020598:Proto-Oncogene Proteins c-myb / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D029681:Arabidopsis Proteins; D020598:Proto-Oncogene Proteins c-myb; D004247:DNA
keywords: Arabidopsis thaliana; DNA binding; LUX ARRHYTHMO; Myb domain; protein–DNA complex