14-3-3 Proteins Form a Guidance Complex with Chloroplast Precursor Proteins in Plants

Correspondence to: Jürgen Soll, jsoll{at}bot.uni-kiel.de (E-mail), 49-431-880-4222 (fax)

Transit sequences of chloroplast-destined precursor proteins are phosphorylated on a serine or threonine residue. The amino acid motif around the phosphorylation site is related to the phosphopeptide binding motif for 14-3-3 proteins. Plant 14-3-3 proteins interact specifically with wheat germ lysate–synthesized chloroplast precursor proteins and require an intact phosphorylation motif within the transit sequence. Chloroplast precursor proteins do not interact with 14-3-3 when synthesized in the heterologous reticulocyte lysate. In contrast, a precursor protein destined for plant mitochondria was found to be associated with 14-3-3 proteins present in the reticulocyte lysate but not with 14-3-3 from wheat germ lysate. This indicates an unrecognized selectivity of 14-3-3 proteins for precursors from mitochondria and plastids in plants in comparison to fungi and animals. The heterooligomeric complex has an apparent size of 200 kD. In addition to the precursor protein, it contains 14-3-3 (probably as a dimer) and a heat shock protein Hsp70 isoform. Dissociation of the precursor complex requires ATP. Protein import experiments of precursor from the oligomeric complex into intact pea chloroplasts reveal three- to fourfold higher translocation rates compared with the free precursor, which is not complexed. We conclude that the 14-3-3–Hsp70–precursor protein complex is a bona fide intermediate in the in vivo protein import pathway in plants.

This article has been cited by other articles:

				M. Ruprecht, T. Bionda, T. Sato, M. S. Sommer, T. Endo, and E. Schleiff On the Impact of Precursor Unfolding during Protein Import into Chloroplasts Mol Plant, January 29, 2010; (2010): ssp116v1 - ssp116. [Abstract] [Full Text] [PDF]

				S. Lee, D. W. Lee, Y. Lee, U. Mayer, Y.-D. Stierhof, S. Lee, G. Jurgens, and I. Hwang Heat Shock Protein Cognate 70-4 and an E3 Ubiquitin Ligase, CHIP, Mediate Plastid-Destined Precursor Degradation through the Ubiquitin-26S Proteasome System in Arabidopsis PLANT CELL, December 1, 2009; 21(12): 3984 - 4001. [Abstract] [Full Text] [PDF]

				A.-K. Berglund, E. Spanning, H. Biverstahl, G. Maddalo, C. Tellgren-Roth, L. Maler, and E. Glaser Dual Targeting to Mitochondria and Chloroplasts: Characterization of Thr-tRNA Synthetase Targeting Peptide Mol Plant, November 1, 2009; 2(6): 1298 - 1309. [Abstract] [Full Text] [PDF]

				D. W. Lee, S. Lee, Y. J. Oh, and I. Hwang Multiple Sequence Motifs in the Rubisco Small Subunit Transit Peptide Independently Contribute to Toc159-Dependent Import of Proteins into Chloroplasts Plant Physiology, September 1, 2009; 151(1): 129 - 141. [Abstract] [Full Text] [PDF]

				H. Inoue and M. Akita Three Sets of Translocation Intermediates Are Formed during the Early Stage of Protein Import into Chloroplasts J. Biol. Chem., March 21, 2008; 283(12): 7491 - 7502. [Abstract] [Full Text] [PDF]

				M. Parsons, A. Karnataki, J. E. Feagin, and A. DeRocher Protein Trafficking to the Apicoplast: Deciphering the Apicomplexan Solution to Secondary Endosymbiosis Eukaryot. Cell, July 1, 2007; 6(7): 1081 - 1088. [Full Text] [PDF]

				A. Schemenewitz, S. Pollmann, C. Reinbothe, and S. Reinbothe A substrate-independent, 14:3:3 protein-mediated plastid import pathway of NADPH:protochlorophyllide oxidoreductase A PNAS, May 15, 2007; 104(20): 8538 - 8543. [Abstract] [Full Text] [PDF]

				B. M. Abell, C. Rabu, P. Leznicki, J. C. Young, and S. High Post-translational integration of tail-anchored proteins is facilitated by defined molecular chaperones J. Cell Sci., May 15, 2007; 120(10): 1743 - 1751. [Abstract] [Full Text] [PDF]

				M. Fan, K. M. Ahmed, M. C. Coleman, D. R. Spitz, and J. J. Li Nuclear Factor-{kappa}B and Manganese Superoxide Dismutase Mediate Adaptive Radioresistance in Low-Dose Irradiated Mouse Skin Epithelial Cells Cancer Res., April 1, 2007; 67(7): 3220 - 3228. [Abstract] [Full Text] [PDF]

				T. Martin, R. Sharma, C. Sippel, K. Waegemann, J. Soll, and U. C. Vothknecht A Protein Kinase Family in Arabidopsis Phosphorylates Chloroplast Precursor Proteins J. Biol. Chem., December 29, 2006; 281(52): 40216 - 40223. [Abstract] [Full Text] [PDF]

				D. G. Durnford and M. W. Gray Analysis of Euglena gracilis Plastid-Targeted Proteins Reveals Different Classes of Transit Sequences Eukaryot. Cell, December 1, 2006; 5(12): 2079 - 2091. [Abstract] [Full Text] [PDF]

				M. Yano, S. Nakamuta, X. Wu, Y. Okumura, and H. Kido A Novel Function of 14-3-3 Protein: 14-3-3{zeta} Is a Heat-Shock-related Molecular Chaperone That Dissolves Thermal-aggregated Proteins Mol. Biol. Cell, November 1, 2006; 17(11): 4769 - 4779. [Abstract] [Full Text] [PDF]

				Y. Xiang, K. Kakani, R. Reade, E. Hui, and D. Rochon A 38-Amino-Acid Sequence Encompassing the Arm Domain of the Cucumber Necrosis Virus Coat Protein Functions as a Chloroplast Transit Peptide in Infected Plants J. Virol., August 15, 2006; 80(16): 7952 - 7964. [Abstract] [Full Text] [PDF]

				S. Li, D. W. Ehrhardt, and S. Y. Rhee Systematic Analysis of Arabidopsis Organelles and a Protein Localization Database for Facilitating Fluorescent Tagging of Full-Length Arabidopsis Proteins Plant Physiology, June 1, 2006; 141(2): 527 - 539. [Abstract] [Full Text] [PDF]

				E.W. Chehab, G. Raman, J.W. Walley, J.V. Perea, G. Banu, S. Theg, and K. Dehesh Rice HYDROPEROXIDE LYASES with Unique Expression Patterns Generate Distinct Aldehyde Signatures in Arabidopsis Plant Physiology, May 1, 2006; 141(1): 121 - 134. [Abstract] [Full Text] [PDF]

				D. W. Lee, S. Lee, G.-j. Lee, K. H. Lee, S. Kim, G.-W. Cheong, and I. Hwang Functional Characterization of Sequence Motifs in the Transit Peptide of Arabidopsis Small Subunit of Rubisco Plant Physiology, February 1, 2006; 140(2): 466 - 483. [Abstract] [Full Text] [PDF]

				J. Bedard and P. Jarvis Recognition and envelope translocation of chloroplast preproteins J. Exp. Bot., September 1, 2005; 56(419): 2287 - 2320. [Abstract] [Full Text] [PDF]

				C. Faul, S. Huttelmaier, J. Oh, V. Hachet, R. H. Singer, and P. Mundel Promotion of importin {alpha}-mediated nuclear import by the phosphorylation-dependent binding of cargo protein to 14-3-3 J. Cell Biol., May 9, 2005; 169(3): 415 - 424. [Abstract] [Full Text] [PDF]

				S. A. Ralph, B. J. Foth, N. Hall, and G. I. McFadden Evolutionary Pressures on Apicoplast Transit Peptides Mol. Biol. Evol., December 1, 2004; 21(12): 2183 - 2194. [Abstract] [Full Text] [PDF]

				J. Yan, C. He, J. Wang, Z. Mao, S. A. Holaday, R. D. Allen, and H. Zhang Overexpression of the Arabidopsis 14-3-3 Protein GF14{lambda} in Cotton Leads to a "Stay-Green" Phenotype and Improves Stress Tolerance under Moderate Drought Conditions Plant Cell Physiol., August 15, 2004; 45(8): 1007 - 1014. [Abstract] [Full Text] [PDF]

				M. D. Smith, C. M. Rounds, F. Wang, K. Chen, M. Afitlhile, and D. J. Schnell atToc159 is a selective transit peptide receptor for the import of nucleus-encoded chloroplast proteins J. Cell Biol., May 10, 2004; 165(3): 323 - 334. [Abstract] [Full Text] [PDF]

				D. M. Bustos and A. A. Iglesias Phosphorylated Non-Phosphorylating Glyceraldehyde-3-Phosphate Dehydrogenase from Heterotrophic Cells of Wheat Interacts with 14-3-3 Proteins Plant Physiology, December 1, 2003; 133(4): 2081 - 2088. [Abstract] [Full Text]

				D. V. Rial, J. Ottado, and E. A. Ceccarelli Precursors with Altered Affinity for Hsp70 in Their Transit Peptides Are Efficiently Imported into Chloroplasts J. Biol. Chem., November 21, 2003; 278(47): 46473 - 46481. [Abstract] [Full Text] [PDF]

				D. Y. Sung and C. L. Guy Physiological and Molecular Assessment of Altered Expression of Hsc70-1 in Arabidopsis. Evidence for Pleiotropic Consequences Plant Physiology, June 1, 2003; 132(2): 979 - 987. [Abstract] [Full Text] [PDF]

				E. Schleiff, M. Jelic, and J. Soll A GTP-driven motor moves proteins across the outer envelope of chloroplasts PNAS, April 15, 2003; 100(8): 4604 - 4609. [Abstract] [Full Text] [PDF]

				S. d. F. Maraschin, G. E. M. Lamers, B. S. de Pater, H. P. Spaink, and M. Wang 14-3-3 isoforms and pattern formation during barley microspore embryogenesis J. Exp. Bot., March 1, 2003; 54(384): 1033 - 1043. [Abstract] [Full Text] [PDF]

				E. Schleiff, J. Soll, M. Kuchler, W. Kuhlbrandt, and R. Harrer Characterization of the translocon of the outer envelope of chloroplasts J. Cell Biol., February 18, 2003; 160(4): 541 - 551. [Abstract] [Full Text] [PDF]

				S. Comparot, G. Lingiah, and T. Martin Function and specificity of 14-3-3 proteins in the regulation of carbohydrate and nitrogen metabolism J. Exp. Bot., January 3, 2003; 54(382): 595 - 604. [Abstract] [Full Text] [PDF]

				C. NDong, J. Danyluk, K. E. Wilson, T. Pocock, N. P.A. Huner, and F. Sarhan Cold-Regulated Cereal Chloroplast Late Embryogenesis Abundant-Like Proteins. Molecular Characterization and Functional Analyses Plant Physiology, July 1, 2002; 129(3): 1368 - 1381. [Abstract] [Full Text] [PDF]

				P. C. Sehnke, J. M. DeLille, and R. J. Ferl Consummating Signal Transduction: The Role of 14-3-3 Proteins in the Completion of Signal-Induced Transitions in Protein Activity PLANT CELL, May 1, 2002; 14(90001): S339 - 354. [Full Text] [PDF]

				H. Fulgosi and J. Soll The Chloroplast Protein Import Receptors Toc34 and Toc159 Are Phosphorylated by Distinct Protein Kinases J. Biol. Chem., March 8, 2002; 277(11): 8934 - 8940. [Abstract] [Full Text] [PDF]

				S. P. Cleary, F.-C. Tan, K.-A. Nakrieko, S. J. Thompson, P. M. Mullineaux, G. P. Creissen, E. von Stedingk, E. Glaser, A. G. Smith, and C. Robinson Isolated Plant Mitochondria Import Chloroplast Precursor Proteins in Vitro with the Same Efficiency as Chloroplasts J. Biol. Chem., February 8, 2002; 277(7): 5562 - 5569. [Abstract] [Full Text] [PDF]

				N. A. Eckardt Transcription Factors Dial 14-3-3 for Nuclear Shuttle PLANT CELL, November 1, 2001; 13(11): 2385 - 2389. [Full Text] [PDF]

				D. Igarashi, S. Ishida, J. Fukazawa, and Y. Takahashi 14-3-3 Proteins Regulate Intracellular Localization of the bZIP Transcriptional Activator RSG PLANT CELL, November 1, 2001; 13(11): 2483 - 2497. [Abstract] [Full Text] [PDF]

				Y. J. Lee, D. H. Kim, Y.-W. Kim, and I. Hwang Identification of a Signal That Distinguishes between the Chloroplast Outer Envelope Membrane and the Endomembrane System in Vivo PLANT CELL, October 1, 2001; 13(10): 2175 - 2190. [Abstract] [Full Text] [PDF]

				T.-S. Yu and H.-m. Li Chloroplast Protein Translocon Components atToc159 and atToc33 Are Not Essential for Chloroplast Biogenesis in Guard Cells and Root Cells Plant Physiology, September 1, 2001; 127(1): 90 - 96. [Abstract] [Full Text] [PDF]

				A. Hiltbrunner, J. Bauer, P.-A. Vidi, S. Infanger, P. Weibel, M. Hohwy, and F. Kessler Targeting of an abundant cytosolic form of the protein import receptor at Toc159 to the outer chloroplast membrane J. Cell Biol., July 23, 2001; 154(2): 309 - 316. [Abstract] [Full Text] [PDF]

				D. Jackson-Constan and K. Keegstra Arabidopsis Genes Encoding Components of the Chloroplastic Protein Import Apparatus Plant Physiology, April 1, 2001; 125(4): 1567 - 1576. [Abstract] [Full Text]

				T. Hirohashi, T. Hase, and M. Nakai Maize Non-Photosynthetic Ferredoxin Precursor Is Mis-Sorted to the Intermembrane Space of Chloroplasts in the Presence of Light Plant Physiology, April 1, 2001; 125(4): 2154 - 2163. [Abstract] [Full Text]

				T. D. Bunney, H. S. van Walraven, and A. H. de Boer 14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase PNAS, March 7, 2001; (2001) 61437498. [Abstract] [Full Text]

				K. Sohrt and J. Soll Toc64, a New Component of the Protein Translocon of Chloroplasts J. Cell Biol., March 20, 2000; 148(6): 1213 - 1222. [Abstract] [Full Text] [PDF]

				T. D. Bunney, H. S. van Walraven, and A. H. de Boer 14-3-3 protein is a regulator of the mitochondrial and chloroplast ATP synthase PNAS, March 27, 2001; 98(7): 4249 - 4254. [Abstract] [Full Text] [PDF]