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Intracellular transport and processing of a tobacco vacuolar β-1,3-glucanase

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Abstract

The class I β-1,3-glucanases are basic, vacuolar enzymes implicated in the defense of plants against pathogen infection. The tobacco (Nicotiana tabacum L.) enzyme is synthesized as a preproprotein with an N-terminal signal peptide for targeting to the lumen of the endoplasmic reticulum and an N-glycosylated C-terminal extension which is lost during protein maturation. The transport and processing of β-1,3-glucanase in cellsuspension cultures of the tobacco cultivar Havana 425 was investigated by pulse-chase labelling and cell fractionation. We verified that mature β-1,3-glucanase is localized in the vacuole of the suspension-cultured cells. Comparison of the time course of processing in homogenates, the soluble fraction, and membrane fractions indicates that proglucanase is transported from the endoplasmic reticulum via the Golgi compartment to the vacuole. Processing to the mature form occurs in the vacuole. Treatment of cells with tunicamycin, which inhibits N-glycosylation, and digestion of the 35S-labelled processing intermediates with endoglycosidase H indicate that β-1,3-glucanase has a single N-glycan attached to the C-terminal extension. Glycosylation is not required for proteolytic processing or correct targeting to the vacuole.

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Abbreviations

endo H:

endoglycosidase H

β-1,3-glucanase:

tobacco class I β-1,3-glucanase

IgG:

immunoglobulin G

Mr :

relative molecular mass

SDS-PAGE:

sodium dodecyl sulfate-polyacrylamide gel electrophoresis

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Author notes

  1. Ursula Hinz

    Present address: Laboratoire de Phytogénétique Cellulaire, Collège Propédeutique, CH-1015, Lausanne, Switzerland

Authors and Affiliations

  1. Friedrich Miescher-Institut, Postfach 2543, CH-4002, Basel, Switzerland

    Liliane Sticher, Ursula Hinz, Alain D. Meyer & Frederick Meins Jr.

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  1. Liliane Sticher
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  2. Ursula Hinz
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  3. Alain D. Meyer
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  4. Frederick Meins Jr.
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Additional information

We thank our colleagues at the Friedrich Miescher Institute Roland Beffa and Arnd Sturm for critical reading of the manuscript and Inge Obergfoell for photography.

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Sticher, L., Hinz, U., Meyer, A.D. et al. Intracellular transport and processing of a tobacco vacuolar β-1,3-glucanase. Planta 188, 559–565 (1992). https://doi.org/10.1007/BF00197049

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  • DOI: https://doi.org/10.1007/BF00197049

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