title: Phosphorylation of Thellungiella salsuginea dehydrins TsDHN-1 and TsDHN-2 facilitates cation-induced conformational changes and actin assembly.
abstract: Group 2 late embryogenesis abundant (LEA) proteins, also known as dehydrins, are intrinsically disordered proteins that are expressed in plants experiencing extreme environmental conditions such as drought or low temperatures. These proteins are characterized by the presence of at least one conserved, lysine-rich K-segment and sometimes by one or more serine-rich S-segments that are phosphorylated. Dehydrins may stabilize proteins and membrane structures during environmental stress and can sequester and scavenge metal ions. Here, we investigate how the conformations of two dehydrins from Thellungiella salsuginea, denoted as TsDHN-1 (acidic) and TsDHN-2 (basic), are affected by pH, interactions with cations and membranes, and phosphorylation. Both TsDHN-1 and TsDHN-2 were expressed as SUMO fusion proteins for in vitro phosphorylation by casein kinase II (CKII), and structural analysis by circular dichroism and attenuated total reflection-Fourier transform infrared spectroscopy. We show that the polyproline II conformation can be induced in the dehydrins by their environmental conditions, including changes in the concentration of divalent cations such as Ca(2+). The assembly of actin by these dehydrins was assessed by sedimentation assays and viewed by transmission electron and atomic force microscopy. Phosphorylation allowed both dehydrins to polymerize actin filaments. These results support the hypothesis that dehydrins stabilize the cytoskeleton under stress conditions and further that phosphorylation may be an important feature of this stabilization.
authors.affiliation: Department of Molecular and Cellular Biology, University of Guelph, Guelph, ON N1G 2W1, Canada.
authors.forename: Luna N
authors.initials: LN
authors.lastname: Rahman
authors.forename: Graham S T
authors.initials: GS
authors.lastname: Smith
authors.forename: Vladimir V
authors.initials: VV
authors.lastname: Bamm
authors.forename: Janine A M
authors.initials: JA
authors.lastname: Voyer-Grant
authors.forename: Barbara A
authors.initials: BA
authors.lastname: Moffatt
authors.forename: John R
authors.initials: JR
authors.lastname: Dutcher
authors.forename: George
authors.initials: G
authors.lastname: Harauz
mesh_terms: D000199:Actins / Q000737:chemistry* / Q000378:metabolism*; D019607:Brassicaceae / Q000737:chemistry / Q000378:metabolism; D002118:Calcium / Q000737:chemistry*; D002413:Cations, Divalent; D006863:Hydrogen-Ion Concentration; D010766:Phosphorylation; D010940:Plant Proteins / Q000737:chemistry* / Q000378:metabolism*; D011487:Protein Conformation; D020033:Protein Isoforms / Q000737:chemistry / Q000378:metabolism; D015032:Zinc / Q000737:chemistry*
chemical_list: D000199:Actins; D002413:Cations, Divalent; D010940:Plant Proteins; D020033:Protein Isoforms; C070556:dehydrin proteins, plant; D015032:Zinc; D002118:Calcium