Abstract
In maturing seed cells, proteins that accumulate in the protein storage vacuoles (PSVs) are synthesized on the endoplasmic reticulum (ER) and transported by vesicles to the PSVs. Vacuolar sorting determinants (VSDs) which are usually amino acid sequences of short or moderate length direct the proteins to this pathway. VSDs identified so far are classified into two types: sequence specific VSDs (ssVSDs) and C-terminal VSDs (ctVSDs). We previously demonstrated that VSDs of α′ and β subunits of β-conglycinin, one of major storage proteins of soybean (Glycine max), reside in the C-terminal ten amino acids. Here we show that both types of VSDs coexist within this region of the α′ subunit. Although ctVSDs can function only at the very C-termini of proteins, the C-terminal ten amino acids of α′ subunit directed green fluorescent protein (GFP) to the PSVs even when they were placed at the N-terminus of GFP, indicating that an ssVSD resides in the sequence. By mutation analysis, it was found that the core sequence of the ssVSD is Ser-Ile-Leu (fifth to seventh residues counted from the C-terminus) which is conserved in the α and β subunits and some vicilin-like proteins. On the other hand, the sequence composed of the C-terminal three amino acids (AFY) directed GFP to the PSVs when it was placed at the C-terminus of GFP, though the function as a VSD was disrupted at the N-terminus of GFP, indicating that the AFY sequence is a ctVSD.
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Acknowledgements
We thank Dr. Y. Niwa (University of Shizuoka) for providing the GFP (S65T) gene. This work was supported in part by grants from the Ministry of Education, Culture, Sports, Science and Technology (to N.M.), and Fuji Foundation for Protein Research (to N.M.).
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Nishizawa, K., Maruyama, N. & Utsumi, S. The C-terminal region of α′ subunit of soybean β-conglycinin contains two types of vacuolar sorting determinants. Plant Mol Biol 62, 111–125 (2006). https://doi.org/10.1007/s11103-006-9007-0
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DOI: https://doi.org/10.1007/s11103-006-9007-0