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Cellular and subcellular distribution of saporins, type-1 ribosome-inactivating proteins, in soapwort (Saponaria officinalis L.)

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Abstract

Many plants contain ribosome-inactivating proteins (RIPs) which are either single enzymatically active polypeptides (type-1 RIPs) or heterodimers (type-2 RIPs) composed of an A-chain, functionally equivalent to a type-1 RIP, which is disulphide bonded to a sugar-binding B-chain. Much attention has focused on the use of RIPs as components of immunotoxins or, more recently, as antiviral agents. In contrast, relatively little is known about either the synthesis and targeting of RIPs or their role within plants. In this study the cellular and subcellular distributions of saporins, the type-1 RIPs from soapwort, have been determined in seeds using immunogold labelling. Saporins were present in the seed storage tissue (perisperm), but are not synthesised in the developing embryo, demonstrating that the expression of saporin genes is subject to tissue-specific control. Within the perisperm, saporin was found in extracellular spaces, in the paramural region between the primary wall and plasmalemma and within the vacuole. In addition, saporin was localised in leaf intercellular spaces. This dual localisation, both vacuolar and extracellular, is significantly different from the localisation of ricin, a type-2 RIP found in castor beans, which is targeted to endosperm protein bodies, and to pokeweed antiviral protein which accumulates in the cell wall matrix of leaf mesophyll cells.

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Abbreviations

PAP:

pokeweed antiviral protein

PBS:

phosphate-buffered saline

RIP:

ribosome-inactivating protein

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Author notes

  1. Anthony P. Fordham-Skelton

    Present address: Department of Biochemistry and Genetics, The Medical School, University of New Castle, NE2 4HH, New Castle, UK

Authors and Affiliations

  1. Department of Biological Sciences, University of Durham, South Road, DH1 3LE, Durham, UK

    Lesley Sinclair, Anthony P. Fordham-Skelton, Nick Harris & Ronald R. D. Croy

  2. Istituto di Patologia Vegetale, Università di Milano, I-Milano, Italy

    Raffaella Carzaniga

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  1. Raffaella Carzaniga
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  2. Lesley Sinclair
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  3. Anthony P. Fordham-Skelton
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Additional information

This work was supported by the UK Agricultural and Food Research Council. R.C. was funded by the Universita di Milano. We would like to thank Martin Hartley and Dorothy Catling for useful discussions and John Gatehouse, Vaughan Hilder and Nigel Robinson for their comments on the manuscript. The authors consider R.C. and L.S. as joint first authors and A.P. F.-S. as the main author for correspondence.

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Carzaniga, R., Sinclair, L., Fordham-Skelton, A.P. et al. Cellular and subcellular distribution of saporins, type-1 ribosome-inactivating proteins, in soapwort (Saponaria officinalis L.). Planta 194, 461–470 (1994). https://doi.org/10.1007/BF00714457

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