An ER-Localized Form of PV72, a Seed-Specific Vacuolar Sorting Receptor, Interferes the Transport of an NPIR-Containing Proteinase in Arabidopsis Leaves

Etsuko Watanabe, Tomoo Shimada, Kentaro Tamura, Ryo Matsushima, Yasuko Koumoto, Mikio Nishimura, Ikuko Hara-Nishimura

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

Putative vacuolar sorting receptors that bind to the vacuolar targeting signals have been found in various plants; pumpkin PV72, pea BP-80 and Arabidopsis AtELP. PV72 is a seed-specific receptor that is predicted to sort seed storage proteins to protein storage vacuoles. Analysis by surface plasmon resonance showed that the lumenal domain of PV72 bound to an NPIR (a typical vacuolar targeting signal)-containing peptide of the precursor of a cysteine proteinase, AtALEU, in the presence of Ca2+ (KD = 0.1 μM). To elucidate the receptor-dependent transport of vacuolar proteins in plant cells, we produced transgenic Arabidopsis plants that expressed a fusion protein (PV72-HDEL) composed of the lumenal domain of PV72 and an endoplasmic reticulum (ER)-retention signal, HDEL. The expression of PV72-HDEL induced the accumulation of the AtALEU precursor. The accumulation level of the AtALEU precursor was dependent on that of PV72-HDEL. In contrast, it did not induce the accumulation of a precursor of another cysteine proteinase, RD21, which contains no NPIR. Detailed subcellular localization revealed that both the AtALEU precursor and PV72-HDEL accumulated in the ER fraction. We found that most of the AtALEU precursor molecules formed a complex with PV72-HDEL. The AtALEU precursor might be trapped by PV72-HDEL in the ER and not transported to the vacuoles. This in-planta analysis supports the hypothesis that an Arabidopsis homolog of PV72 functions as a sorting receptor for the NPIR-containing proteinase. The overall results suggest that vacuolar sorting receptors for the protein storage vacuoles and the lytic vacuoles share the similar recognition mechanism for a vacuolar targeting signal.

Original languageEnglish
Pages (from-to)9-17
Number of pages9
JournalPlant and Cell Physiology
Volume45
Issue number1
DOIs
Publication statusPublished - Jan 2004
Externally publishedYes

Fingerprint

Vacuoles
Arabidopsis
Endoplasmic Reticulum
endoplasmic reticulum
sorting
Seeds
Peptide Hydrolases
proteinases
vacuoles
receptors
Cysteine Proteases
seeds
cysteine proteinases
leaves
storage proteins
Seed Storage Proteins
Cucurbita
Surface Plasmon Resonance
Genetically Modified Plants
Peas

Keywords

  • Arabidopsis thaliana
  • Ligand-receptor interaction
  • PV72
  • Vacuolar proteins
  • Vacuolar sorting receptor
  • Vacuolar targeting signal

ASJC Scopus subject areas

  • Plant Science
  • Physiology
  • Cell Biology

Cite this

An ER-Localized Form of PV72, a Seed-Specific Vacuolar Sorting Receptor, Interferes the Transport of an NPIR-Containing Proteinase in Arabidopsis Leaves. / Watanabe, Etsuko; Shimada, Tomoo; Tamura, Kentaro; Matsushima, Ryo; Koumoto, Yasuko; Nishimura, Mikio; Hara-Nishimura, Ikuko.

In: Plant and Cell Physiology, Vol. 45, No. 1, 01.2004, p. 9-17.

Research output: Contribution to journalArticle

Watanabe, Etsuko ; Shimada, Tomoo ; Tamura, Kentaro ; Matsushima, Ryo ; Koumoto, Yasuko ; Nishimura, Mikio ; Hara-Nishimura, Ikuko. / An ER-Localized Form of PV72, a Seed-Specific Vacuolar Sorting Receptor, Interferes the Transport of an NPIR-Containing Proteinase in Arabidopsis Leaves. In: Plant and Cell Physiology. 2004 ; Vol. 45, No. 1. pp. 9-17.
@article{efa15b9fbeb64049997273738831e852,
title = "An ER-Localized Form of PV72, a Seed-Specific Vacuolar Sorting Receptor, Interferes the Transport of an NPIR-Containing Proteinase in Arabidopsis Leaves",
abstract = "Putative vacuolar sorting receptors that bind to the vacuolar targeting signals have been found in various plants; pumpkin PV72, pea BP-80 and Arabidopsis AtELP. PV72 is a seed-specific receptor that is predicted to sort seed storage proteins to protein storage vacuoles. Analysis by surface plasmon resonance showed that the lumenal domain of PV72 bound to an NPIR (a typical vacuolar targeting signal)-containing peptide of the precursor of a cysteine proteinase, AtALEU, in the presence of Ca2+ (KD = 0.1 μM). To elucidate the receptor-dependent transport of vacuolar proteins in plant cells, we produced transgenic Arabidopsis plants that expressed a fusion protein (PV72-HDEL) composed of the lumenal domain of PV72 and an endoplasmic reticulum (ER)-retention signal, HDEL. The expression of PV72-HDEL induced the accumulation of the AtALEU precursor. The accumulation level of the AtALEU precursor was dependent on that of PV72-HDEL. In contrast, it did not induce the accumulation of a precursor of another cysteine proteinase, RD21, which contains no NPIR. Detailed subcellular localization revealed that both the AtALEU precursor and PV72-HDEL accumulated in the ER fraction. We found that most of the AtALEU precursor molecules formed a complex with PV72-HDEL. The AtALEU precursor might be trapped by PV72-HDEL in the ER and not transported to the vacuoles. This in-planta analysis supports the hypothesis that an Arabidopsis homolog of PV72 functions as a sorting receptor for the NPIR-containing proteinase. The overall results suggest that vacuolar sorting receptors for the protein storage vacuoles and the lytic vacuoles share the similar recognition mechanism for a vacuolar targeting signal.",
keywords = "Arabidopsis thaliana, Ligand-receptor interaction, PV72, Vacuolar proteins, Vacuolar sorting receptor, Vacuolar targeting signal",
author = "Etsuko Watanabe and Tomoo Shimada and Kentaro Tamura and Ryo Matsushima and Yasuko Koumoto and Mikio Nishimura and Ikuko Hara-Nishimura",
year = "2004",
month = "1",
doi = "10.1093/pcp/pch012",
language = "English",
volume = "45",
pages = "9--17",
journal = "Plant and Cell Physiology",
issn = "0032-0781",
publisher = "Oxford University Press",
number = "1",

}

TY - JOUR

T1 - An ER-Localized Form of PV72, a Seed-Specific Vacuolar Sorting Receptor, Interferes the Transport of an NPIR-Containing Proteinase in Arabidopsis Leaves

AU - Watanabe, Etsuko

AU - Shimada, Tomoo

AU - Tamura, Kentaro

AU - Matsushima, Ryo

AU - Koumoto, Yasuko

AU - Nishimura, Mikio

AU - Hara-Nishimura, Ikuko

PY - 2004/1

Y1 - 2004/1

N2 - Putative vacuolar sorting receptors that bind to the vacuolar targeting signals have been found in various plants; pumpkin PV72, pea BP-80 and Arabidopsis AtELP. PV72 is a seed-specific receptor that is predicted to sort seed storage proteins to protein storage vacuoles. Analysis by surface plasmon resonance showed that the lumenal domain of PV72 bound to an NPIR (a typical vacuolar targeting signal)-containing peptide of the precursor of a cysteine proteinase, AtALEU, in the presence of Ca2+ (KD = 0.1 μM). To elucidate the receptor-dependent transport of vacuolar proteins in plant cells, we produced transgenic Arabidopsis plants that expressed a fusion protein (PV72-HDEL) composed of the lumenal domain of PV72 and an endoplasmic reticulum (ER)-retention signal, HDEL. The expression of PV72-HDEL induced the accumulation of the AtALEU precursor. The accumulation level of the AtALEU precursor was dependent on that of PV72-HDEL. In contrast, it did not induce the accumulation of a precursor of another cysteine proteinase, RD21, which contains no NPIR. Detailed subcellular localization revealed that both the AtALEU precursor and PV72-HDEL accumulated in the ER fraction. We found that most of the AtALEU precursor molecules formed a complex with PV72-HDEL. The AtALEU precursor might be trapped by PV72-HDEL in the ER and not transported to the vacuoles. This in-planta analysis supports the hypothesis that an Arabidopsis homolog of PV72 functions as a sorting receptor for the NPIR-containing proteinase. The overall results suggest that vacuolar sorting receptors for the protein storage vacuoles and the lytic vacuoles share the similar recognition mechanism for a vacuolar targeting signal.

AB - Putative vacuolar sorting receptors that bind to the vacuolar targeting signals have been found in various plants; pumpkin PV72, pea BP-80 and Arabidopsis AtELP. PV72 is a seed-specific receptor that is predicted to sort seed storage proteins to protein storage vacuoles. Analysis by surface plasmon resonance showed that the lumenal domain of PV72 bound to an NPIR (a typical vacuolar targeting signal)-containing peptide of the precursor of a cysteine proteinase, AtALEU, in the presence of Ca2+ (KD = 0.1 μM). To elucidate the receptor-dependent transport of vacuolar proteins in plant cells, we produced transgenic Arabidopsis plants that expressed a fusion protein (PV72-HDEL) composed of the lumenal domain of PV72 and an endoplasmic reticulum (ER)-retention signal, HDEL. The expression of PV72-HDEL induced the accumulation of the AtALEU precursor. The accumulation level of the AtALEU precursor was dependent on that of PV72-HDEL. In contrast, it did not induce the accumulation of a precursor of another cysteine proteinase, RD21, which contains no NPIR. Detailed subcellular localization revealed that both the AtALEU precursor and PV72-HDEL accumulated in the ER fraction. We found that most of the AtALEU precursor molecules formed a complex with PV72-HDEL. The AtALEU precursor might be trapped by PV72-HDEL in the ER and not transported to the vacuoles. This in-planta analysis supports the hypothesis that an Arabidopsis homolog of PV72 functions as a sorting receptor for the NPIR-containing proteinase. The overall results suggest that vacuolar sorting receptors for the protein storage vacuoles and the lytic vacuoles share the similar recognition mechanism for a vacuolar targeting signal.

KW - Arabidopsis thaliana

KW - Ligand-receptor interaction

KW - PV72

KW - Vacuolar proteins

KW - Vacuolar sorting receptor

KW - Vacuolar targeting signal

UR - http://www.scopus.com/inward/record.url?scp=1042279798&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1042279798&partnerID=8YFLogxK

U2 - 10.1093/pcp/pch012

DO - 10.1093/pcp/pch012

M3 - Article

C2 - 14749481

AN - SCOPUS:1042279798

VL - 45

SP - 9

EP - 17

JO - Plant and Cell Physiology

JF - Plant and Cell Physiology

SN - 0032-0781

IS - 1

ER -