title: Arabidopsis dynamin-related proteins, DRP2A and DRP2B, function coordinately in post-Golgi trafficking.
abstract: Dynamin-related proteins (DRPs) are large GTPases involved in a wide range of cellular membrane remodeling processes. In Arabidopsis thaliana, two paralogous land plant-specific type DRPs, DRP2A and DRP2B, are thought to participate in the regulation of post-Golgi trafficking. Here, we examined their molecular properties and functional relationships. qRT-PCR and GUS assays showed that DRP2A and DRP2B were expressed ubiquitously, although their expressions were strongest around root apical meristems and vascular bundles. Yeast two-hybrid, bi-molecular fluorescent complementation, and co-immunoprecipitation mass spectrometry analyses revealed that DRP2A and DRP2B interacted with each other. In observations with confocal laser scanning microscopy and variable incidence angle fluorescent microscopy, fluorescent fusions of DRP2A and DRP2B almost completely co-localized and were mainly localized to endocytic vesicle formation sites of the plasma membrane, clathrin-enriched trans-Golgi network and the cell plate in root epidermal cells. Treatments with wortmannin, an inhibitor of phosphatidylinositol 3-/4-kinases, latrunculin B, an inhibitor of actin polymerization, and oryzalin, an inhibitor of microtubule polymerization, increased the resident time of DRP2A and DRP2B on the plasma membrane. These results show that DRP2A and DRP2B function coordinately in multiple pathways of post-Golgi trafficking in phosphatidylinositol 3- or 4-kinase and cytoskeleton polymerization-dependent manners.
authors.affiliation: Laboratory of Plant Molecular Genetics, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Jiahe
authors.initials: J
authors.lastname: Huang
authors.affiliation: Laboratory of Plant Molecular Genetics, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan.
authors.forename: Masaru
authors.initials: M
authors.lastname: Fujimoto
authors.affiliation: Plant Global Education Project, Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Masayuki
authors.initials: M
authors.lastname: Fujiwara
authors.affiliation: Plant Global Education Project, Graduate School of Biological Sciences, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0192, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Fukao
authors.affiliation: Laboratory of Plant Molecular Genetics, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Japan Science and Technology Agency (JST), PRESTO, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan.
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Arimura
authors.affiliation: Laboratory of Plant Molecular Genetics, Graduate School of Agricultural and Life Sciences, The University of Tokyo, 1-1-1 Yayoi, Bunkyo-ku, Tokyo 113-8657, Japan; Japan Science and Technology Agency (JST), CREST, 4-1-8 Honcho, Kawaguchi, Saitama 332-0012, Japan. Electronic address: atsutsu@mail.ecc.u-tokyo.ac.jp.
authors.forename: Nobuhiro
authors.initials: N
authors.lastname: Tsutsumi
mesh_terms: D000199:Actins / Q000378:metabolism; D000730:Androstadienes / Q000737:chemistry; D017360:Arabidopsis / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D019086:Bridged Bicyclo Compounds, Heterocyclic / Q000737:chemistry; D002462:Cell Membrane / Q000378:metabolism; D003599:Cytoskeleton / Q000378:metabolism; D004136:Dinitrobenzenes / Q000737:chemistry; D034281:Dynamins / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D019204:GTP-Binding Proteins / Q000378:metabolism*; D006056:Golgi Apparatus / Q000378:metabolism*; D013058:Mass Spectrometry; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D018517:Plant Roots / Q000378:metabolism; D021381:Protein Transport; D013424:Sulfanilamides / Q000737:chemistry; D053778:Thiazolidines / Q000737:chemistry; D020798:Two-Hybrid System Techniques; D000077191:Wortmannin
chemical_list: D000199:Actins; D000730:Androstadienes; D029681:Arabidopsis Proteins; D019086:Bridged Bicyclo Compounds, Heterocyclic; D004136:Dinitrobenzenes; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D013424:Sulfanilamides; D053778:Thiazolidines; C012465:oryzalin; C434467:ADL6 protein, Arabidopsis; C539085:DRP2B protein, Arabidopsis; D019204:GTP-Binding Proteins; D034281:Dynamins; C037068:latrunculin B; D000077191:Wortmannin
keywords: Arabidopsis; DRP2; Dynamin-related protein; Post-Golgi trafficking