title: Differential Regulation of Clathrin and Its Adaptor Proteins during Membrane Recruitment for Endocytosis.
abstract: In plants, clathrin-mediated endocytosis (CME) is dependent on the function of clathrin and its accessory heterooligomeric adaptor protein complexes, ADAPTOR PROTEIN2 (AP-2) and the TPLATE complex (TPC), and is negatively regulated by the hormones auxin and salicylic acid (SA). The details for how clathrin and its adaptor complexes are recruited to the plasma membrane (PM) to regulate CME, however, are poorly understood. We found that SA and the pharmacological CME inhibitor tyrphostin A23 reduce the membrane association of clathrin and AP-2, but not that of the TPC, whereas auxin solely affected clathrin membrane association, in Arabidopsis (Arabidopsis thaliana). Genetic and pharmacological experiments revealed that loss of AP2μ or AP2σ partially affected the membrane association of other AP-2 subunits and that the AP-2 subunit AP2σ, but not AP2μ, was required for SA- and tyrphostin A23-dependent inhibition of CME Furthermore, we show that although AP-2 and the TPC are both required for the PM recruitment of clathrin in wild-type cells, the TPC is necessary for clathrin PM association in AP-2-deficient cells. These results indicate that developmental signals may differentially modulate the membrane recruitment of clathrin and its core accessory complexes to regulate the process of CME in plant cells.
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Chao
authors.initials: C
authors.lastname: Wang
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Tianwei
authors.initials: T
authors.lastname: Hu
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Xu
authors.initials: X
authors.lastname: Yan
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Tingting
authors.initials: T
authors.lastname: Meng
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Yutong
authors.initials: Y
authors.lastname: Wang
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Qingmei
authors.initials: Q
authors.lastname: Wang
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Xiaoyue
authors.initials: X
authors.lastname: Zhang
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Ying
authors.initials: Y
authors.lastname: Gu
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Clara
authors.initials: C
authors.lastname: Sánchez-Rodríguez
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Astrid
authors.initials: A
authors.lastname: Gadeyne
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Jinxing
authors.initials: J
authors.lastname: Lin
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Staffan
authors.identifier: 0000-0002-6377-5132
authors.initials: S
authors.lastname: Persson
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Daniël
authors.identifier: 0000-0002-9385-4851
authors.initials: D
authors.lastname: Van Damme
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Chuanyou
authors.identifier: 0000-0003-0202-3890
authors.initials: C
authors.lastname: Li
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.).
authors.forename: Sebastian Y
authors.identifier: 0000-0001-7465-1787
authors.initials: SY
authors.lastname: Bednarek
authors.affiliation: College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China (C.W., T.H., X.Y., T.M., Y.W., Q.W., J.P.);State Key Laboratory of Plant Genomics, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, Beijing 100101, China (X.Z., C.L.);Department of Biochemistry and Molecular Biology, Pennsylvania State University, University Park, Pennsylvania 16802 (Y.G.);Department of Biology, Institute of Agricultural Sciences, Eidgenössisch Technische Hochschule Zurich, 8092 Zurich, Switzerland (C.S.-R.);Department of Plant Systems Biology, Vlaams Instituut voor Biotechnologie, Department of Plant Biotechnology and Bioinformatics, Ghent University, B-9052 Ghent, Belgium (A.G., D.V.D.);College of Biological Sciences and Biotechnology, Beijing Forestry University, Beijing 100083, China (J.L.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Biosciences, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Biochemistry, University of Wisconsin, Madison, Wisconsin 53706 (S.Y.B.) jwpan@zjnu.cn.
authors.forename: Jianwei
authors.identifier: 0000-0001-5155-4933
authors.initials: J
authors.lastname: Pan
mesh_terms: D033962:Adaptor Protein Complex 2 / Q000187:drug effects / Q000235:genetics / Q000378:metabolism*; D017360:Arabidopsis / Q000166:cytology* / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002966:Clathrin / Q000187:drug effects / Q000378:metabolism*; D033922:Clathrin Heavy Chains / Q000187:drug effects / Q000378:metabolism; D033941:Clathrin Light Chains / Q000187:drug effects / Q000378:metabolism; D022163:Clathrin-Coated Vesicles / Q000187:drug effects / Q000378:metabolism; D004705:Endocytosis / Q000502:physiology*; D006112:Gravitation; D007210:Indoleacetic Acids / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism; D008566:Membranes / Q000378:metabolism*; D009154:Mutation; D018517:Plant Roots / Q000378:metabolism; D030821:Plants, Genetically Modified; D020156:Salicylic Acid / Q000378:metabolism / Q000494:pharmacology; D050656:Transcription Factor AP-2 / Q000378:metabolism; D020032:Tyrphostins / Q000037:antagonists & inhibitors
chemical_list: D033962:Adaptor Protein Complex 2; D029681:Arabidopsis Proteins; D002966:Clathrin; D033941:Clathrin Light Chains; D007210:Indoleacetic Acids; D008565:Membrane Proteins; C518499:TPLATE protein, Arabidopsis; D050656:Transcription Factor AP-2; D020032:Tyrphostins; D033922:Clathrin Heavy Chains; D020156:Salicylic Acid; C060641:tyrphostin A23