title: Transport and sorting of the solanum tuberosum sucrose transporter SUT1 is affected by posttranslational modification.
abstract: The plant sucrose transporter SUT1 from Solanum tuberosum revealed a dramatic redox-dependent increase in sucrose transport activity when heterologously expressed in Saccharomyces cerevisiae. Plant plasma membrane vesicles do not show any change in proton flux across the plasma membrane in the presence of redox reagents, indicating a SUT1-specific effect of redox reagents. Redox-dependent sucrose transport activity was confirmed electrophysiologically in Xenopus laevis oocytes with SUT1 from maize (Zea mays). Localization studies of green fluorescent protein fusion constructs showed that an oxidative environment increased the targeting of SUT1 to the plasma membrane where the protein concentrates in 200- to 300-nm raft-like microdomains. Using plant plasma membranes, St SUT1 can be detected in the detergent-resistant membrane fraction. Importantly, in yeast and in plants, oxidative reagents induced a shift in the monomer to dimer equilibrium of the St SUT1 protein and increased the fraction of dimer. Biochemical methods confirmed the capacity of SUT1 to form a dimer in plants and yeast cells in a redox-dependent manner. Blue native PAGE, chemical cross-linking, and immunoprecipitation, as well as the analysis of transgenic plants with reduced expression of St SUT1, confirmed the dimerization of St SUT1 and Sl SUT1 (from Solanum lycopersicum) in planta. The ability to form homodimers in plant cells was analyzed by the split yellow fluorescent protein technique in transiently transformed tobacco (Nicotiana tabacum) leaves and protoplasts. Oligomerization seems to be cell type specific since under native-like conditions, a phloem-specific reduction of the dimeric form of the St SUT1 protein was detectable in SUT1 antisense plants, whereas constitutively inhibited antisense plants showed reduction only of the monomeric form. The role of redox control of sucrose transport in plants is discussed.
authors.affiliation: Institute of Biology, Department of Plant Physiology, Humboldt University, 10115 Berlin, Germany.
authors.forename: Undine
authors.initials: U
authors.lastname: Krügel
authors.forename: Liesbeth M
authors.initials: LM
authors.lastname: Veenhoff
authors.forename: Jennifer
authors.initials: J
authors.lastname: Langbein
authors.forename: Elena
authors.initials: E
authors.lastname: Wiederhold
authors.forename: Johannes
authors.initials: J
authors.lastname: Liesche
authors.forename: Thomas
authors.initials: T
authors.lastname: Friedrich
authors.forename: Bernhard
authors.initials: B
authors.lastname: Grimm
authors.forename: Enrico
authors.initials: E
authors.lastname: Martinoia
authors.forename: Bert
authors.initials: B
authors.lastname: Poolman
authors.forename: Christina
authors.initials: C
authors.lastname: Kühn
mesh_terms: D000818:Animals; D004594:Electrophysiology; D005260:Female; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D047468:Immunoprecipitation; D008564:Membrane Potentials; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D018613:Microscopy, Confocal; D009865:Oocytes / Q000378:metabolism / Q000502:physiology; D010084:Oxidation-Reduction; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology; D011499:Protein Processing, Post-Translational; D021381:Protein Transport; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D011198:Solanum tuberosum / Q000235:genetics / Q000378:metabolism*; D013395:Sucrose / Q000378:metabolism; D014982:Xenopus laevis
chemical_list: D026901:Membrane Transport Proteins; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; C078729:sucrose transport protein, plant; D049452:Green Fluorescent Proteins; D013395:Sucrose
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