title: CESA TRAFFICKING INHIBITOR inhibits cellulose deposition and interferes with the trafficking of cellulose synthase complexes and their associated proteins KORRIGAN1 and POM2/CELLULOSE SYNTHASE INTERACTIVE PROTEIN1.
abstract: Cellulose synthase complexes (CSCs) at the plasma membrane (PM) are aligned with cortical microtubules (MTs) and direct the biosynthesis of cellulose. The mechanism of the interaction between CSCs and MTs, and the cellular determinants that control the delivery of CSCs at the PM, are not yet well understood. We identified a unique small molecule, CESA TRAFFICKING INHIBITOR (CESTRIN), which reduces cellulose content and alters the anisotropic growth of Arabidopsis (Arabidopsis thaliana) hypocotyls. We monitored the distribution and mobility of fluorescently labeled cellulose synthases (CESAs) in live Arabidopsis cells under chemical exposure to characterize their subcellular effects. CESTRIN reduces the velocity of PM CSCs and causes their accumulation in the cell cortex. The CSC-associated proteins KORRIGAN1 (KOR1) and POM2/CELLULOSE SYNTHASE INTERACTIVE PROTEIN1 (CSI1) were differentially affected by CESTRIN treatment, indicating different forms of association with the PM CSCs. KOR1 accumulated in bodies similar to CESA; however, POM2/CSI1 dissociated into the cytoplasm. In addition, MT stability was altered without direct inhibition of MT polymerization, suggesting a feedback mechanism caused by cellulose interference. The selectivity of CESTRIN was assessed using a variety of subcellular markers for which no morphological effect was observed. The association of CESAs with vesicles decorated by the trans-Golgi network-localized protein SYNTAXIN OF PLANTS61 (SYP61) was increased under CESTRIN treatment, implicating SYP61 compartments in CESA trafficking. The properties of CESTRIN compared with known CESA inhibitors afford unique avenues to study and understand the mechanism under which PM-associated CSCs are maintained and interact with MTs and to dissect their trafficking routes in etiolated hypocotyls.
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Natasha
authors.identifier: http://orcid.org/0000-0002-0804-2884
authors.initials: N
authors.lastname: Worden
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Wilkop
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Victor Esteva
authors.initials: VE
authors.lastname: Esteve
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Richard
authors.initials: R
authors.lastname: Jeannotte
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Rahul
authors.initials: R
authors.lastname: Lathe
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Samantha
authors.initials: S
authors.lastname: Vernhettes
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Bart
authors.initials: B
authors.lastname: Weimer
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Glenn
authors.initials: G
authors.lastname: Hicks
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Jose
authors.initials: J
authors.lastname: Alonso
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: John
authors.initials: J
authors.lastname: Labavitch
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: Staffan
authors.initials: S
authors.lastname: Persson
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.).
authors.forename: David
authors.initials: D
authors.lastname: Ehrhardt
authors.affiliation: Departments of Plant Sciences (N.W., T.E.W., V.E.E., J.L., G.D.) and Veterinary Medicine (R.J., B.W.), University of California, Davis, California 95616;Max-Planck-Institute of Molecular Plant Physiology, Science Campus, 14476 Golm, Germany (R.L., S.P.);Institut National de la Recherche Agronomique, Institute Jean-Pierre Bourgin, 78026 Versailles, France (S.V.);Department of Botany and Plant Sciences, University of California, Riverside, California 92521 (G.H.);Department of Plant and Microbial Biology, North Caroline State University, Raleigh, North Carolina 27695 (J.A.);Australian Research Council Centre of Excellence in Plant Cell Walls, School of Botany, University of Melbourne, Parkville, Victoria 3010, Australia (S.P.); andDepartment of Plant Biology, Carnegie Institution for Science, Stanford, California 94305 (D.E.) gdrakakaki@ucdavis.edu.
authors.forename: Georgia
authors.initials: G
authors.lastname: Drakakaki
mesh_terms: D016880:Anisotropy; D017360:Arabidopsis / Q000166:cytology / Q000201:enzymology / Q000254:growth & development / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000378:metabolism*; D001549:Benzamides / Q000494:pharmacology; D002352:Carrier Proteins / Q000378:metabolism*; D002451:Cell Compartmentation / Q000187:drug effects; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D002480:Cellulase / Q000378:metabolism*; D002482:Cellulose / Q000378:metabolism*; D004136:Dinitrobenzenes / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D005947:Glucose / Q000378:metabolism; D005964:Glucosyltransferases / Q000378:metabolism*; D049452:Green Fluorescent Proteins / Q000378:metabolism; D018546:Hypocotyl / Q000187:drug effects / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism; D046912:Multiprotein Complexes / Q000378:metabolism*; D021381:Protein Transport / Q000187:drug effects; D013424:Sulfanilamides / Q000494:pharmacology
chemical_list: D029681:Arabidopsis Proteins; D001549:Benzamides; C551801:CSI1 protein, Arabidopsis; D002352:Carrier Proteins; D004136:Dinitrobenzenes; D004791:Enzyme Inhibitors; C429844:KOR1 protein, Arabidopsis; D008565:Membrane Proteins; D046912:Multiprotein Complexes; D013424:Sulfanilamides; D049452:Green Fluorescent Proteins; C012465:oryzalin; C057467:isoxaben; D002482:Cellulose; D005964:Glucosyltransferases; C478648:cellulose synthase; D002480:Cellulase; D005947:Glucose