title: ASG2 is a farnesylated DWD protein that acts as ABA negative regulator in Arabidopsis.
abstract: The tagging-via-substrate approach designed for the capture of mammal prenylated proteins was adapted to Arabidopsis cell culture. In this way, proteins are in vivo tagged with an azide-modified farnesyl moiety and captured thanks to biotin alkyne Click-iT® chemistry with further streptavidin-affinity chromatography. Mass spectrometry analyses identified four small GTPases and ASG2 (ALTERED SEED GERMINATION 2), a protein previously associated to the seed germination gene network. ASG2 is a conserved protein in plants and displays a unique feature that associates WD40 domains and tetratricopeptide repeats. Additionally, we show that ASG2 has a C-terminal CaaX-box that is farnesylated in vitro. Protoplast transfections using CaaX prenyltransferase mutants show that farnesylation provokes ASG2 nucleus exclusion. Moreover, ASG2 interacts with DDB1 (DAMAGE DNA BINDING protein 1), and the subcellular localization of this complex depends on ASG2 farnesylation status. Finally, germination and root elongation experiments reveal that asg2 and the farnesyltransferase mutant era1 (ENHANCED RESPONSE TO ABSCISIC ACID (ABA) 1) behave in similar manners when exposed to ABA or salt stress. To our knowledge, ASG2 is the first farnesylated DWD (DDB1 binding WD40) protein related to ABA response in Arabidopsis that may be linked to era1 phenotypes.
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', UFR des Sciences et Techniques, Université François Rabelais de Tours, Tours, F-37200, France.
authors.forename: Christelle
authors.initials: C
authors.lastname: Dutilleul
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', UFR des Sciences et Techniques, Université François Rabelais de Tours, Tours, F-37200, France.
authors.forename: Iliana
authors.initials: I
authors.lastname: Ribeiro
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', UFR des Sciences et Techniques, Université François Rabelais de Tours, Tours, F-37200, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Blanc
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, 06520-8104, USA.
authors.forename: Cynthia D
authors.initials: CD
authors.lastname: Nezames
authors.affiliation: Department of Molecular, Cellular and Developmental Biology, Yale University, New Haven, CT, 06520-8104, USA.
authors.forename: Xing Wang
authors.initials: XW
authors.lastname: Deng
authors.affiliation: Department of Plant Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, 30-387, Poland.
authors.forename: Piotr
authors.initials: P
authors.lastname: Zglobicki
authors.affiliation: Universidad de Antioquia, Laboratorio de Biotecnología, Sede de Investigación Universitaria, Medellín, Colombia.
authors.forename: Ana María
authors.initials: AM
authors.lastname: Palacio Barrera
authors.affiliation: Universidad de Antioquia, Laboratorio de Biotecnología, Sede de Investigación Universitaria, Medellín, Colombia.
authors.forename: Lucia
authors.initials: L
authors.lastname: Atehortùa
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', UFR des Sciences et Techniques, Université François Rabelais de Tours, Tours, F-37200, France.
authors.forename: Martine
authors.initials: M
authors.lastname: Courtois
authors.affiliation: INRA, UMR85 Physiologie de la Reproduction et des Comportements, UMR CNRS 7247, UFR, IFC, Plate-forme d'Analyse Intégrative des Biomolécules, Laboratoire de Spectrométrie de Masse, Nouzilly, F-37380, France.
authors.forename: Valérie
authors.initials: V
authors.lastname: Labas
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', UFR des Sciences et Techniques, Université François Rabelais de Tours, Tours, F-37200, France.
authors.forename: Nathalie
authors.initials: N
authors.lastname: Giglioli-Guivarc'h
authors.affiliation: EA2106 'Biomolécules et Biotechnologies Végétales', UFR des Sciences et Techniques, Université François Rabelais de Tours, Tours, F-37200, France.
authors.forename: Eric
authors.initials: E
authors.lastname: Ducos
mesh_terms: D000040:Abscisic Acid / Q000378:metabolism*; D000595:Amino Acid Sequence; D017360:Arabidopsis / Q000166:cytology / Q000187:drug effects / Q000235:genetics* / Q000502:physiology; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D018525:Germination; D008969:Molecular Sequence Data; D009154:Mutation; D010641:Phenotype; D010937:Plant Growth Regulators / Q000378:metabolism*; D018517:Plant Roots / Q000166:cytology / Q000187:drug effects / Q000235:genetics / Q000502:physiology; D030821:Plants, Genetically Modified; D017368:Protein Prenylation; D021381:Protein Transport; D011993:Recombinant Fusion Proteins; D016415:Sequence Alignment; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological; D015293:Transducin / Q000235:genetics / Q000378:metabolism*
chemical_list: C000611520:ASG2 protein, Arabidopsis; D029681:Arabidopsis Proteins; D010937:Plant Growth Regulators; D011993:Recombinant Fusion Proteins; D012965:Sodium Chloride; D000040:Abscisic Acid; D015293:Transducin
keywords: ABA signalling; DDB1 CUL4 E3 ubiquitin ligase; protein farnesylation; tagging-via-substrate