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BiP and zein binding domains within the delta zein protein

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Abstract

Zeins are alcohol soluble seed storage proteins synthesized within the endosperm of maize and subsequently deposited into endoplasmic reticulum (ER) derived protein bodies. The genes encoding the beta and delta zeins were previously introduced into tobacco with the expectation of improving the nutritional quality of plants (Bagga et al. in Plant Physiol 107:13, 1997). Novel protein bodies are produced in the leaves of transgenic plants accumulating the beta or delta zein proteins. The mechanism of protein body formation within leaves is unknown. It is also unknown how zeins are retained in the ER since they do not contain known ER retention motifs. Retention may be due to an interaction of zeins with an ER chaperone such as binding luminal protein (BiP). We have demonstrated protein–protein interactions with the delta zeins, beta zeins, and BiP proteins using an E. coli two-hybrid system. In this study, four putative BiP binding motifs were identified within the delta zein protein using a BiP scoring program (Blond-Elguindi et al. in Cell 75:717, 1993). These putative binding motifs were mutated and their effects on protein interactions were analyzed in both a prokaryotic two-hybrid system and in plants. These mutations resulted in reduced BiP–zein protein interaction and also altered zein–zein interactions. Our results indicate that specific motifs are necessary for BiP–delta zein protein interactions and that there are specific motifs which are necessary for zein–zein interactions. Furthermore, our data demonstrates that zein proteins must be able to interact with BiP and zeins for their stability and ability to form protein bodies.

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Abbreviations

BiP:

Binding luminal protein

ER:

Endoplasmic reticulum

References

  • Bagga S, Adams H, Kemp JD, Sengupta-Gopalan C (1995) Accumulation of 15-kilodalton zein in novel protein bodies in transgenic tobacco. Plant Physiol 107:13–23

    CAS  PubMed  Google Scholar 

  • Bagga S, Adams HP, Rodriguez FD, Kemp JD, Sengupta-Gopalan C (1997) Coexpression of the maize delta-zein and beta-zein genes results in stable accumulation of delta-zein in endoplasmic reticulum-derived protein bodies formed by beta-zein. Plant Cell 9:1683–96

    Google Scholar 

  • Bellucci M, Lazzari B, Viotti A, Arcioni S (1997) Differential expression of g-zein gene in Medicago sativa, Lotus corniculatus and Nicotiana tabacum. Plant Sci 127:161–169

    Google Scholar 

  • Bellucci M, Alpini A, Paolocci F, Cong L, Arcioni S (2000) Accumulation of maize g-zein and g-zein:KDEL to high levels in tobacco leaves and differential increase of BiP synthesis in transformants. Theor Appl Genet 101:796–804

    Google Scholar 

  • Blond-Elguindi S, Cwirla SE, Dower WJ, Lipshutz RJ, Sprang SR, Sambrook JF, Gething MJ (1993) Affinity panning of a library of peptides displayed on bacteriophages reveals the binding specificity of BiP. Cell 75:717–728

    Article  CAS  PubMed  Google Scholar 

  • Boston RS, Fontes EB, Shank BB, Wrobel RL (1991) Increased expression of the maize immunoglobulin binding protein homolog b-70 in three zein regulatory mutants. Plant Cell 3:497–505

    Google Scholar 

  • Coleman CE, Herman EM, Takasaki K, Larkins BA (1996) The maize gamma-zein sequesters alpha-zein and stabilizes its accumulation in protein bodies of transgenic tobacco endosperm. Plant Cell 8:2335–2345

    Google Scholar 

  • Ems-McClung SC, Benmoussa M, Hainline BE (2002) Mutational analysis of the maize gamma zein C-terminal cysteine residues. Plant Sci 2002:131–141

    Google Scholar 

  • Foresti O, Frigerio L, Holkeri H, deVirgilio M, Vavassori S, Vitale A (2003) A phaseolin domain involved directly in trimer assembly is a determinant for binding by the chaperone BiP. The Plant Cell 15:2464–2475

    Google Scholar 

  • Geli MI, Torrent M, Ludevid D (1994) Two structural domains mediate two sequential events in [gamma]-zein targeting: protein endoplasmic reticulum retention and protein body formation. Plant Cell 6:1911–1922

    Google Scholar 

  • Geourjon C, Deleage G (1994) SOPM: a self optimized prediction method for protein secondary structure prediction. Protein Eng 7:157–164

    Google Scholar 

  • Ghoshroy S, Citovsky V (1998) Preservation of plant cell ultrastructure during immunolocalization of virus particles. J Virol Methods 74:223–229

    Google Scholar 

  • Hinchliffe DJ, Kemp JD (2002) b-Zein protein bodies sequester and protect the 18-kDa d-zein protein from degradation. Plant Sci 163:741–752

    Google Scholar 

  • Horton R (1989) Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene 77:61–68

    Google Scholar 

  • Kim CS, Woo Ym YM, Clore AM, Burnett RJ, Carneiro NP, Larkins BA (2002) Zein protein interactions, rather than the asymmetric distribution of zein mRNAs on endoplasmic reticulum membranes, influence protein body formation in maize endosperm. Plant Cell 14:655–72

    Google Scholar 

  • Kirihara JA, Hunsperger JP, Mahoney WC, Messing JW (1988) Differential expression of a gene for a methionine-rich storage protein in maize. Mol Gen Genet 211:477–484

    Google Scholar 

  • Knarr G, Modrow S, Todd A, Gething MJ, Buchnert J (1999) BiP-binding sequences in HIV gp160. J Biol Chem 274:29850–29857

    Google Scholar 

  • Kogan, MJ, Lopez O, Cocera M, Lopez-Iglesias C, De La Maza A, Giralt E (2004) Exploring the interaction of the surfactant N-terminal domain of gamma-zein with soybean phosphatidylcholine liposomes. Biopolymers 73(2):258–268

    Google Scholar 

  • Leborgne-Castel N, Jelitto-Van Dooren EPWM, Crofts AR, Denecke J (1999) Overexpression of BiP in tobacco alleviates endoplasmic reticulum stress. Plant Cell 11:459–469

    Google Scholar 

  • Lending CR, Larkins BA (1989) Changes in the zein composition of protein bodies during maize endosperm development. Plant Cell 1:1011–1023

    Google Scholar 

  • Li X, Wu Y, Zhang DZ, Gillikin JW, Boston RS, Franceschi VR, Okita TW (1993) Rice prolamine protein body biogenesis: a BiP mediated Process

    Google Scholar 

  • Mainieri D, Rossi M, Archiniti M, Bellucci M, De Marchis F, Vavassori S, Pompa A, Arcioni S, Vitale A (2004) Zeolin. A new recombinant storage protein constructed using maize gamma-zein and bean phaseolin. Plant Physiol 136:3447–3456

    Google Scholar 

  • Molinari M, Galli C, Piccaluga V, Pieren M, Paganetti P (2002) Sequential assistance of molecular chaperones and transient formation of covalent complexes during protein degradation from the ER. J Cell Biol 158:247–257

    Google Scholar 

  • Ng DTW, Spear ED, Walter P (2000) The Unfolded Protein Response regulates multiple apsects of secretory and membrane protein biogenesis and endoplasmic reticulum quality control. J Cell Biol 150:77–88

    Google Scholar 

  • Nuttall J, Vitale A, Frigerio L (2003) C-terminal extension of phaseolin with a short methionine-rich sequence can inhibit trimerisation and result in high instability. Plant Mol Biol 51:885–894

    Google Scholar 

  • Randall J (1997) Targeting and stabilization of 10 kD and 15 kD zeins in transgenic tobacco. NMSU

  • Randall J, Bagga S, Adams H, Kemp JD (2000) A modified 10-kDa zein protein produces two morphologically distinct protein bodies in transgenic tobacco. Plant Sci 150:21–28

    Google Scholar 

  • Randall J, Sutton D, Ghoshroy S, Bagga S, Kemp JD (2004) Co-ordinate expression of b- and d-zeins in transgenic tobacco. Plant Sci 167:267–372

    Google Scholar 

  • Reynolds A, Lundbland V, Dorris D, Keaveney M (1997) Assay for b-galactosidase in liquid culture. Current protocols in molecular biology. Wiley, New York, pp 13.6.1–13.6.6

    Google Scholar 

  • Rogers S, Klee H, Horsch R, Fraley R (1987) Improved vectors for plant transformation: expression cassette vectors and new identifiable markers. Meth Enzymol 153:253–263

    Google Scholar 

  • Tottey S, Rondet SAM, Borrelly G, Robinson PJ, Rich PR, Robinson NJ (2002) A copper metallochaperone for photosynthesis and respiration reveals metal-specific targets, interaction with an importer, and alternative sites for copper acquisition. J Biol Chem 277:5490–5497

    Google Scholar 

  • Van Regenmortal MHV (2002) A paradigm shift is needed in proteomics: structure determines function should be replaced by binding determines function. J Mol Recognit 15:349–351

    Google Scholar 

  • Vitale A, Ceriotti A (2004) Protein Quality control mechanisms and protein storage in the endoplasmic reticulum. A conflict of interests?. Plant Physiol 136:3420–3426

    Google Scholar 

  • Voinnet O, Rivas S, Mestre P, Baulcombe D (2003) An enhanced transient expression system in plants based on suppression of gene silencing by the p19 protein of tomato bushy stunt virus. Plant J 33:949–956

    Google Scholar 

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Acknowledgments

The authors would like to thank Jennifer Jackson for her superb technical support of this work. The authors also thank Lorenzo Aleman for the actin primers and his critical reading of the manuscript. The authors express their gratitude to Dr. Soumitra Ghoshroy for his assistance with the EM work. The authors thank Dr. Suman Bagga for the providing the pMEZ construct, Dr. Doug Hinchliffe for providing the pMH316-18 construct, and Dr. Jose Louis Ortoga for the oliga dT primer. The authors would also like to thank Dr. Sylvie Blond-Elguindi for the BiP program, Dr. Denecke for the blp4 gene, Dr. Boston for the BiP polyclonal antibodies, and Dr. Messing for the δ zein polyclonal antibodies. The research was supported by special grant 2003-34250-13279.

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Authors and Affiliations

  1. Department of Entomology, Plant Pathology, and Weed Science, New Mexico State University, 3BE Skeen Hall, Las Cruces, NM, 88003, USA

    Jennifer J Randall, Dennis W Sutton, Stephen F. Hanson & John D. Kemp

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  1. Jennifer J Randall
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  2. Dennis W Sutton
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  3. Stephen F. Hanson
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  4. John D. Kemp
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Correspondence to Jennifer J Randall or John D. Kemp.

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Randall, J.J., Sutton, D.W., Hanson, S.F. et al. BiP and zein binding domains within the delta zein protein. Planta 221, 656–666 (2005). https://doi.org/10.1007/s00425-005-1482-z

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  • DOI: https://doi.org/10.1007/s00425-005-1482-z

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