title: Sporopollenin biosynthetic enzymes interact and constitute a metabolon localized to the endoplasmic reticulum of tapetum cells.
abstract: The sporopollenin polymer is the major constituent of exine, the outer pollen wall. Recently fatty acid derivatives have been shown to be the precursors of sporopollenin building units. ACYL-COA SYNTHETASE, POLYKETIDE SYNTHASE A (PKSA) and PKSB, TETRAKETIDE α-PYRONE REDUCTASE1 (TKPR1) and TKPR2 have been demonstrated to be involved in sporopollenin biosynthesis in Arabidopsis (Arabidopsis thaliana). Here all these sporopollenin biosynthetic enzymes but TKPR2 have been immunolocalized to endoplasmic reticulum of anther tapetal cells. Pull-down experiments demonstrated that tagged recombinant proteins interacted to form complexes whose constituents were characterized by immunoblotting. In vivo protein interactions were evidenced by yeast (Saccharomyces cerevisiae) two-hybrid analysis and by fluorescence lifetime imaging microscopy/Förster resonance energy transfer studies in transgenic Nicotiana benthamiana, which were used to test the possibility that the enzymes interact to form a biosynthetic metabolon. Various pairs of proteins fused to two distinct fluorochromes were coexpressed in N. benthamiana leaf tissues and fluorescence lifetime imaging microscopy/Förster resonance energy transfer measurements demonstrated that proteins interacted pairwise in planta. Taken together, these results suggest the existence of a sporopollenin metabolon.
authors: Institut de Biologie Moléculaire des Plantes, Unité Propre de Recherche 2357 du Centre National de la Recherche Scientifique, Université de Strasbourg, 67084 Strasbourg cedex, France.
authors: Benjamin
authors: B
authors: Lallemand
authors: Mathieu
authors: M
authors: Erhardt
authors: Thierry
authors: T
authors: Heitz
authors: Michel
authors: M
authors: Legrand
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism*; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D001704:Biopolymers / Q000096:biosynthesis* / Q000235:genetics; D002338:Carotenoids / Q000096:biosynthesis* / Q000235:genetics; D003066:Coenzyme A Ligases / Q000235:genetics / Q000378:metabolism; D004721:Endoplasmic Reticulum / Q000235:genetics / Q000378:metabolism*; D004798:Enzymes / Q000235:genetics / Q000378:metabolism; D035264:Flowers / Q000235:genetics / Q000378:metabolism; D031541:Fluorescence Resonance Energy Transfer; D030821:Plants, Genetically Modified; D048630:Polyketide Synthases / Q000235:genetics / Q000378:metabolism*; D011994:Recombinant Proteins / Q000235:genetics / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014026:Nicotiana / Q000235:genetics; D020798:Two-Hybrid System Techniques
chemical_list: D029681:Arabidopsis Proteins; D001704:Biopolymers; D004798:Enzymes; C552540:LAP5 protein, Arabidopsis; C552541:LAP6 protein, Arabidopsis; D011994:Recombinant Proteins; C009800:sporopollenin; D002338:Carotenoids; D048630:Polyketide Synthases; C542001:ACOS5 protein, Arabidopsis; D003066:Coenzyme A Ligases
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