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Large alkyl side-chains of isoleucine and leucine in the NPIRL region constitute the core of the vacuolar sorting determinant of sporamin precursor

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Abstract

The N-terminal propeptide of the sporamin precursor contains vacuolar targeting information within the Asn-26/Pro-27/Ile-28/Arg-29/Leu-30 (NPIRL) sequence. An Agrobacterium-mediated transient expression assay with tobacco BY-2 cells was employed to investigate the role of each amino acid of the NPIRL region in vacuolar targeting. Replacement of Asn-26, Pro-27, Ile-28 and Leu-30 with several amino acids caused secretion of the mutant prosporamin. Leu was the only amino acid that could be substituted for Ile-28 without affecting transport. Exchange of Leu-30 for amino acids with small side-chains abolished vacuolar delivery. These results indicate that the consensus composition of the NPIRL sequence is [preferably Asn]-[not acidic]-[Ile or Leu]-[any amino acid]-[large and hydrophobic] and suggest that the large alkyl side-chains of Ile-28 and Leu-30 constitute the core of the vacuolar sorting determinant.

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References

  • Ahmed, S.U., Bar-Peled, M. and Raikhel, N.V. 1997. Cloning and subcellular location of an Arabidopsis receptor-like protein that shares common features with protein-sorting receptors of eukaryotic cells. Plant Physiol. 114: 325–336.

    PubMed  Google Scholar 

  • Bednarek, S.Y. and Raikhel, N.V. 1991. The barley lectin carboxyl-terminal propeptide is a vacuolar protein sorting determinant in plants. Plant Cell 3: 1195–1206.

    Article  PubMed  Google Scholar 

  • Di Sansebastiano, G.-P, Paris, N., Marc-Martin, S. and Neuhaus, J-M. 1998. Specific accumulation of GFP in a non-acidic vac-uolar compartment via a C-terminal propeptide-mediated sorting pathway. Plant J. 15: 449–457.

    PubMed  Google Scholar 

  • Dombrowski, J.E., Schroeder, M.R., Bednarek, S.Y. and Raikhel, N.V. 1993. Determination of the functional elements within the vacuolar targeting signal of barley lectin. Plant Cell 5: 587–596.

    PubMed  Google Scholar 

  • Frigerio, L., de Virgilio, M., Prada, A., Faoro, F. and Vitale, A. 1998. Sorting of phaseolin to the vacuole is saturable and requires a short C-terminal peptide. Plant Cell 10: 1031–1042.

    PubMed  Google Scholar 

  • Gomord, V., Denmat, L., Fitchette-Lainé, A.C., Satiat-Jeunemaitre, B., Hawes, C. and Faye, L. 1997. The C-terminal HDEL se-quence is sufficient for retention of secretory proteins in the endoplasmic reticulum (ER) but promotes vacuolar targeting of proteins that escape the ER. Plant J. 11: 313–325.

    PubMed  Google Scholar 

  • Hattori, T., Ichihara, S. and Nakamura, K. 1987. Processing of a plant vacuolar protein precursor in vitro. Eur. J. Biochem. 166: 533–538.

    PubMed  Google Scholar 

  • Hattori, T., Yoshida, N. and Nakamura, K. 1989. Structural relation-ship among the members of a multigene family coding for the sweet potato tuberous root storage protein. Plant Mol. Biol. 13: 563–572.

    PubMed  Google Scholar 

  • Hinz, G., Hillmer, S., Bäumer, M. and Hohl, I. 1999. Vacuolar storage proteins and the putative vacuolar sorting receptor BP-80 exit the Golgi apparatus of developing pea cotyledons in different transport vesicles. Plant Cell 11: 1509–1524.

    PubMed  Google Scholar 

  • Holwerda, B.C., Padgett, H.S. and Rogers, J.C. 1992. Efficient vac-uolar targeting of proaleurain is mediated by interaction of short contiguous peptide determinants. Plant Cell 4: 307–318.

    Google Scholar 

  • Hong, E., Davidson, A.R. and Kaiser, C.A. 1996. A pathway for targeting soluble misfolded proteins to the yeast vacuole. J. Cell Biol. 135: 623–633.

    PubMed  Google Scholar 

  • Ishikawa, A., Ohta, S., Matsuoka, K., Hattori, T. and Naka-mura, K. 1994. A family of potato genes that encode Kunitz-type proteinase inhibitors: structural comparison and differential expression. Plant Cell Physiol. 35: 303–312.

    PubMed  Google Scholar 

  • Kirsch T., Paris N., Butler J.M., Beevers L., and Rogers J.C. 1994 Purification and initial characterization of a potential plant vacuolar targeting receptor. Proc. Natl. Acad. Sci. USA 91: 3403–3407.

    PubMed  Google Scholar 

  • Kirsch T., Saalbach G., Raikhel N.V., and Beevers L. 1996. Interac-tion of a potential vacuolar targeting receptor with amino-and carboxyl-terminal targeting determinants. Plant Physiol. 111: 469–474.

    PubMed  Google Scholar 

  • Koide, Y., Hirano, H., Matsuoka, K. and Nakamura, K. 1997. The N-terminal propeptide of the precursor to sporamin acts as a vacuole-targeting signal even at the C terminus of the mature part in tobacco cells. Plant Physiol. 114: 863–870.

    PubMed  Google Scholar 

  • Koide, Y., Matsuoka, K., Ohto, M-a, and Nakamura, K. 1999. The N-terminal propeptide and the C-terminus of the precursor to 20 kilo-dalton potato tuber protein can function as different types of vacuolar sorting signal. Plant Cell Physiol. 40: 1152–1159.

    PubMed  Google Scholar 

  • Matsuoka, K. and Bednarek, S.Y. 1998. Protein transport within the plant cell endomembrane system: an update. Curr. Opin. Plant Biol. 1: 463–469.

    PubMed  Google Scholar 

  • Matsuoka, K. and Nakamura, K. 1991. Propeptide of a precursor to a plant vacuolar protein required for vacuolar targeting. Proc. Natl. Acad. Sci. USA 88: 834–838.

    PubMed  Google Scholar 

  • Matsuoka, K. and Neuhaus, J.-M. 1999. Cis-element of targeting to the vacuole. J. Exp. Bot. 50: 165–174.

    Google Scholar 

  • Matsuoka, K., Matsumoto, S., Hattori, T., Machida, S. and Nakamura, K. 1990. Vacuolar targeting and post-translational processing of the precursors to the sweet potato tuberous root storage protein in heterologous plant cells. J. Biol. Chem. 265: 19750–19757.

    PubMed  Google Scholar 

  • Matsuoka, K., Bassham, D.C., Raikhel, N.V. and Nakamura, K. 1995a. Different sensitivity to wortmannin of two vacuolar sort-ing signals indicates the presence of distinct sorting machineries in tobacco cells. J. Cell Biol. 6: 1307–1318.

    Google Scholar 

  • Matsuoka, K., Watanabe, N. and Nakamura, K. 1995b. O-glycosylation of a precursor to a sweet potato vacuolar protein, sporamin, expressed in tobacco cells. Plant J. 8: 877–889.

    PubMed  Google Scholar 

  • Matsuoka, K., Higuchi, T., Maeshima, M. and Nakamura, K. 1997. A vacuolar-type H C-ATPase in a nonvacuolar organelle is re-quired for the sorting of soluble vacuolar protein precursors in tobacco cells. Plant Cell 9: 533–535.

    PubMed  Google Scholar 

  • Melchers, L.S., Sela-Buurlage, M.B., Vloemans, S.A., Woloshuk, C.P., van Roekel, J.S.C., Pen, J., van den Elzen, P.J.M. and Cornelissen, B.J.C. 1993. Extracellular targeting of the vacuo-lar tobacco proteins AP24, chitinase and _-1,3-glucanase in transgenic plants. Plant Mol. Biol. 21: 583–593.

    PubMed  Google Scholar 

  • Miller, E.H., Lee, M.C.S. and Anderson, M.A. 1999. Identification and characterization of a prevacuolar compartment in stigmas of Nicotiana alata. Plant Cell 11: 1499–1508.

    PubMed  Google Scholar 

  • Nakamura, K. and Matsuoka, K. 1993. Protein targeting to the vacuole in plant cells. Plant Physiol. 101: 1–5.

    PubMed  Google Scholar 

  • Nakamura, K., Matsuoka, K., Mukumoto, F. and Watanabe, N. 1993. Processing and transport to the vacuole of a precursor to sweet potato sporamin in transformed tobacco cell line BY-2. J. Exp. Bot. 44 (Suppl): 331–338.

    Google Scholar 

  • Neuhaus, J.-M. and Rogers, J.C. 1998. Sorting of proteins to vacuoles in plant cells. Plant Mol. Biol. 38: 127–144.

    PubMed  Google Scholar 

  • Neuhaus, J.-M., Sticher, L., Meins F, Jr. and Boller, T. 1991. A short C-terminal sequence is necessary and sufficient for the targeting of chitinases to the plant vacuole. Proc. Natl. Acad. Sci. USA 88: 10362–10366.

    PubMed  Google Scholar 

  • Neuhaus, J.-M., Pietrzak, M. and Boller, T. 1994. Mutational analysis of the C-terminal vacuolar targeting peptide of tobacco chitinase: low specificity of the sorting system, and gradual transition between intracellular retention and secretion into the extracellular space. Plant J. 5: 45–54.

    PubMed  Google Scholar 

  • Ohta, S., Mita, S., Hattori, T. and Nakamura, K. 1990. Construction and expression in tobacco of a _-glucuronidase (GUS) reporter gene containing an intron within the coding sequence. Plant Cell Physiol. 31: 805–813.

    Google Scholar 

  • Okamoto, T. and Minamikawa, T. 1999. Molecular cloning and characterization of Vigna mungo processing enzyme 1 (VmPE-1), an asparaginyl endopeptidase possibly involved in post-translational processing of a vacuolar cysteine endopeptidase (SH-EP). Plant Mol. Biol. 39: 63–73.

    PubMed  Google Scholar 

  • Okita, T.W. and Rogers, J.C. 1996. Compartmentalization of pro-teins in the endomembrane system of plant cells. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47: 327–350.

    PubMed  Google Scholar 

  • Paris, N., Stanley, C.M., Jones, R.L. and Rogers, J.C. 1996. Plant cells contain two functionally distinct vacuolar compartments. Cell 85: 563–572.

    PubMed  Google Scholar 

  • Paris, N., Rogers, S.W., Jiang, L., Kirsch, T., Beevers, L., Phillips, T.E. and Rogers, J.C. 1997. Molecular cloning and further char-acterization of a probable plant vacuolar sorting receptor. Plant Physiol. 115: 29–39.

    PubMed  Google Scholar 

  • Robinson, D.G., Bäumer, M, Hinz, G. and Hohl, I. 1998. Vesicle transfer of storage proteins to the vacuole: the role of the Golgi apparatus and multivesicular bodies. J. Plant Physiol. 152: 659–667.

    Google Scholar 

  • Saalbach, G., Jung, R., Kunze, G., Saalbach, I., Adler, K. and Müntz, K. 1991. Different legumin protein domains act as vacuolar targeting signals. Plant Cell 3: 695–708.

    Article  PubMed  Google Scholar 

  • Saalbach, G., Rosso, M. and Schumann, U. 1996. The vacuolar targeting signal of the 2S albumin from Brazil nut resides at the C terminus and involves the C-terminal propeptide as an essential element. Plant Physiol. 112: 975–985.

    PubMed  Google Scholar 

  • Sanderfoot, A.A., Ahmed, S.U., Marty-Mazars, D., Rapoport, I., Kirchhausen, T., Marty, F. and Raikhel, N.V. 1998. A putative vacuolar cargo receptor partially colocalizes with AtPEP12p on a prevacuolar compartment in Arabidopsis roots. Proc. Natl. Acad. Sci. USA 95: 9920–9925.

    PubMed  Google Scholar 

  • Schroeder, M.R., Borkhsenious, O.N., Matsuoka, K., Nakamura, K. and Raikhel, N.V. 1993. Colocalization of barley lectin and sporamin in vacuoles of transgenic tobacco plants. Plant Physiol 101: 451–458.

    PubMed  Google Scholar 

  • Shimada, T., Kuroyanagi, M., Nishimura, M. and Hara-Nishimura, I. 1997. A pumpkin 72-kDa membrane protein of precursor-accumulating vesicles has characteristics of a vacuolar sorting receptor. Plant Cell Physiol. 38: 1414–1420.

    PubMed  Google Scholar 

  • van Voorst, F., Kielland-Brandt, M.C. and Winther, J.R. 1996. Mutational analysis of the vacuolar sorting signal of procar-boxypeptidase Y in yeast shows a low requirement for sequence conservation. J. Biol. Chem. 271: 841–8460.

    PubMed  Google Scholar 

  • von Schaewen, A. and Chrispeels, M.J. 1993. Identification of vac-uolar sorting information in phytohemagglutinin, an unprocessed vacuolar protein. J. Exp. Bot. 44 (Suppl): 339–342.

    Google Scholar 

  • Watanabe, H., Abe, K., Emori, Y., Hosoyama, H. and Arai, S. 1991. Molecular cloning and gibberellin-induced expression of multi-ple cysteine proteinases of rice seeds (oryzains). J. Biol. Chem. 266: 16897–16902.

    PubMed  Google Scholar 

  • Yoshioka, Y., Takahashi, Y., Matsuoka, K., Nakamura, K., Koizumi, J., Kojima, M. and Machida, Y. 1996. Transient gene expression in plant cells mediated by Agrobacterium tumefaciens: applica-tion for the analysis of virulence loci. Plant Cell Physiol. 37: 782–789.

    Google Scholar 

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  1. Laboratory of Biochemistry, Graduate School of Bioagricultural Sciences, Nagoya University, Chikusa-ku, Nagoya, 464-8601, Japan

    Ken Matsuoka & Kenzo Nakamura

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  1. Ken Matsuoka
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  2. Kenzo Nakamura
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Matsuoka, K., Nakamura, K. Large alkyl side-chains of isoleucine and leucine in the NPIRL region constitute the core of the vacuolar sorting determinant of sporamin precursor. Plant Mol Biol 41, 825–835 (1999). https://doi.org/10.1023/A:1006357413084

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