title: Toc33 and Toc64-III cooperate in precursor protein import into the chloroplasts of Arabidopsis thaliana.
abstract: The import of cytosolically synthesized precursor proteins into chloroplasts by the translocon at the outer envelope membrane of chloroplasts (TOC) is crucial for organelle function. The recognition of precursor proteins at the chloroplast surface precedes translocation and involves the membrane-inserted receptor subunits Toc34 and Toc159. A third receptor, Toc64, was discussed to recognize cytosolic complexes guiding precursor proteins to the membrane surface, but this function remains debated. We analysed Arabidopsis thaliana plants carrying a T-DNA insertion in the gene encoding the Toc64 homolog Toc64-III. We observed a light intensity-dependent growth phenotype, which is distinct from the phenotype of ppi1, the previously described mutant of the TOC34 homolog TOC33. Furthermore, chloroplast import of the model precursor proteins pOE33 and pSSU into chloroplasts is reduced in protoplasts isolated from plants with impaired Toc64-III function. This suggests that Toc64-III modulates the translocation efficiency in vivo. A ppi1 and toc64-III double mutant shows a significant increase in the transcript levels of HSP90 and TOC75-III, the latter coding for the pore-forming TOC component. Remarkably, the protein level of Toc75-III is significantly reduced, suggesting that Toc64-III and Toc33 cooperate in the insertion or stabilization of Toc75-III. Accordingly, the results presented support Toc64 as an import-relevant component of the TOC complex.
authors.affiliation: Center of Membrane Proteomics, Cluster of Excellence Macromolecular Complexes Frankfurt, Department of Biosciences, Molecular Cell Biology of Plants, Johann-Wolfgang-Goethe University Frankfurt, D-60438 Frankfurt, Germany.
authors.forename: Manuel
authors.initials: M
authors.lastname: Sommer
authors.forename: Mareike
authors.initials: M
authors.lastname: Rudolf
authors.forename: Bodo
authors.initials: B
authors.lastname: Tillmann
authors.forename: Joanna
authors.initials: J
authors.lastname: Tripp
authors.forename: Maik S
authors.initials: MS
authors.lastname: Sommer
authors.forename: Enrico
authors.initials: E
authors.lastname: Schleiff
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000378:metabolism* / Q000528:radiation effects; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism; D002736:Chloroplasts / Q000235:genetics / Q000378:metabolism*; D003600:Cytosol / Q000378:metabolism; D004269:DNA, Bacterial / Q000378:metabolism; D055786:Gene Knockout Techniques; D007425:Intracellular Membranes / Q000378:metabolism; D008027:Light; D008565:Membrane Proteins / Q000235:genetics / Q000378:metabolism*; D010641:Phenotype; D010788:Photosynthesis; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism / Q000528:radiation effects; D025941:Protein Interaction Mapping; D021381:Protein Transport; D011523:Protoplasts / Q000378:metabolism; D013312:Stress, Physiological
chemical_list: D029681:Arabidopsis Proteins; D002352:Carrier Proteins; D004269:DNA, Bacterial; D008565:Membrane Proteins; C469717:Ppi1 protein, Arabidopsis; C036483:T-DNA; C114786:Toc33 protein, Arabidopsis; C488374:Toc64 protein, Arabidopsis