title: PIN2 turnover in Arabidopsis root epidermal cells explored by the photoconvertible protein Dendra2.
abstract: The steady state level of integral membrane proteins is dependent on a strictly controlled delivery and removal. Here we show that Dendra2, a green-to-red photoconvertible fluorescent protein, is a suitable tool to study protein turnover in plants. We characterized the fluorescence properties of Dendra2 expressed either as a free protein or as a tag in Arabidopsis thaliana roots and optimized photoconversion settings to study protein turnover. Dendra2 was fused to the PIN2 protein, an auxin transporter in the root tip, and by time-lapse imaging and assessment of red and green signal intensities in the membrane after photoconversion we quantified directly and simultaneously the rate of PIN2 delivery of the newly synthesized protein into the plasma membrane as well as the disappearance of the protein from the plasma membrane due to degradation. Additionally we have verified several factors which are expected to affect PIN2 protein turnover and therefore potentially regulate root growth.
authors: Max Planck Institute for Plant Breeding Research, Köln, Germany. jan.jasik@yahoo.com
authors: Ján
authors: J
authors: Jásik
authors: Barbara
authors: B
authors: Boggetti
authors: František
authors: F
authors: Baluška
authors: Dieter
authors: D
authors: Volkmann
authors: Thomas
authors: T
authors: Gensch
authors: Twan
authors: T
authors: Rutten
authors: Thomas
authors: T
authors: Altmann
authors: Elmon
authors: E
authors: Schmelzer
mesh_terms: D000040:Abscisic Acid / Q000494:pharmacology; D000693:Anaerobiosis; D017360:Arabidopsis / Q000502:physiology*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D003080:Cold Temperature; D003513:Cycloheximide / Q000494:pharmacology; D003517:Cyclopentanes / Q000494:pharmacology; D003609:Dactinomycin / Q000494:pharmacology; D003624:Darkness; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D008164:Luminescent Proteins / Q000378:metabolism* / Q000528:radiation effects; D018613:Microscopy, Confocal; D054883:Oxylipins / Q000494:pharmacology; D018517:Plant Roots / Q000378:metabolism*; D030821:Plants, Genetically Modified; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D000040:Abscisic Acid; D029681:Arabidopsis Proteins; D003513:Cycloheximide; D003517:Cyclopentanes; D003609:Dactinomycin; D004121:Dimethyl Sulfoxide; D008164:Luminescent Proteins; D054883:Oxylipins; D011993:Recombinant Fusion Proteins; C583169:Dendra2 protein, Dendronephthya; C117013:AT5G57090 protein, Arabidopsis; C011006:jasmonic acid
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