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Deposition of a recombinant peptide in ER-derived protein bodies by retention with cysteine-rich prolamins in transgenic rice seed

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Abstract

A 7Crp peptide composed of seven major human T cell epitopes derived from the Japanese cedar pollen allergens Cry j 1 and Cry j 2 is an ideal tolerogen for peptide immunotherapy against Japanese cedar pollinosis. To maximize the accumulation level of the 7Crp peptide in transgenic rice seed, we tested endosperm specific promoters and intracellular localizations suitable for stable accumulation. A 7Crp peptide carrying the KDEL ER retention signal directed by the 2.3-kb promoter of the glutelin GluB-1, which contains a signal peptide, accumulated at the highest level of about 60 μg/grain. Notably, the 7Crp peptide predominantly accumulated in ER-derived protein bodies irrespective of the presence of various sorting signals or expression as a fusion protein with glutelin. We attribute this abnormal pattern of accumulation to the formation of disulfide bonds between the 7Crp peptide and cysteine-rich (Cys-rich) prolamin storage proteins. Furthermore, the formation of these aggregates induced the chaperone proteins BiP and PDI as an ER stress response.

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Abbreviations

AGPase:

ADP-glucose pyrophosphorylase

GluB-1:

Rice glutelin B1

GluB-4:

Rice glutelin B4

GluA-2:

Rice glutelin A2

Tg:

Transgenic

BiP:

Binding protein

PDI:

Protein disulfide isomerase

PSV:

Protein storage vacuole

PB-I/II:

Protein body type-I/II

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Acknowledgments

We thank Dr. S. Saito (Jikei University School Medicine) for the gift of mouse monocolonal 7Crp antibody. This work was supported by the research grant, “Functional analysis of genes relevant to Agriculturally important traits in the rice genome IP2001” and “Genomics for Agricultural Innovation GMC0009” from the Ministry of Agriculture, Forestry and Fisheries of Japan to F. Takaiwa.

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Authors and Affiliations

  1. Transgenic Crop Research and Development Center, National Institute of Agrobiological Sciences, Kannondai 2-1-2, Tsukuba, Ibaraki, 305-8602, Japan

    Fumio Takaiwa, Sakiko Hirose, Hidenori Takagi, Lijun Yang & Yuhya Wakasa

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  1. Fumio Takaiwa
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  2. Sakiko Hirose
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  3. Hidenori Takagi
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  4. Lijun Yang
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  5. Yuhya Wakasa
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Correspondence to Fumio Takaiwa.

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Takaiwa, F., Hirose, S., Takagi, H. et al. Deposition of a recombinant peptide in ER-derived protein bodies by retention with cysteine-rich prolamins in transgenic rice seed. Planta 229, 1147–1158 (2009). https://doi.org/10.1007/s00425-009-0905-7

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