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Expression of the ubiquitin variant ubR48 decreases proteolytic activity in Arabidopsis and induces cell death

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Abstract

The ubiquitin–proteasome pathway is the major route for protein degradation in eukaryotes. We show here that this pathway can be inhibited in Arabidopsis thaliana by expression of a ubiquitin variant that contains Arg instead of Lys at position 48 (ubR48). A major consequence of ubR48 expression is the induction of cell death. Cell death induction coincides with the appearance of reactive oxygen intermediates, but is independent of salicylic acid. We found changes in expression of some defense-related genes, but these changes are apparently insufficient to cause alterations in the response to a bacterial pathogen. Expression of ubR48 from an inducible gene allowed investigation of kinetic parameters of cell death induction. In the absence of additional stress factors, slow death processes dominate if the transgene is induced in seedlings older than 2 weeks. The inducible gene also allowed isolation of suppressor mutants. Expression of ubR48 may cause changes similar to inhibition of the proteasome, which also induces various forms of cell death. Thus, ubR48 is a tool to manipulate protein turnover and to probe cell death programs in plants.

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Abbreviations

CaMV:

Cauliflower mosaic virus

Dex:

Dexamethasone

DHFR:

Dihydrofolate reductase

GUS:

β-glucuronidase from E. coli

HR:

Hypersensitive response

Me-JA:

Jasmonic acid–methyl ester

MTX:

Methotrexate

NBT:

Nitroblue tetrazolium

PCD:

Programmed cell death

ROI:

Reactive oxygen intermediate

SA:

Salicylic acid

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Acknowledgements

We wish to thank J. Celenza (Boston University, Boston, USA), J. Dangl, and M. A. Torres (University of North Carolina, Chapel Hill, USA) and Syngenta Biotechnology Inc. (Research Triangle Park, USA) for plants and bacteria, S. Stary (Univ. of Vienna, Vienna, Austria) for sharing materials, F. Hadacek for help with unpublished experiments, and M.-L. Kalda for photography. This work was supported by the Austrian Science Foundation FWF (grant P 13927), and by the Max Planck Society.

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  1. Peter Schlögelhofer & Claudia Kerzendorfer

    Present address: Department of Chromosome Biology, Vienna Biocenter 2, University of Vienna, Dr. Bohr-Gasse 1, 1030, Vienna, Austria

  2. Viktoria Nizhynska

    Present address: Center for Brain Research, Medical University of Vienna, Spitalgasse 4, 1090, Vienna, Austria

Authors and Affiliations

  1. Institute of Botany, University of Vienna, Rennweg 14, 1030, Vienna, Austria

    Peter Schlögelhofer, Claudia Kerzendorfer, Viktoria Nizhynska & Andreas Bachmair

  2. Department of Plant Developmental Biology, Max Planck Institute for Plant Breeding Research, Carl-von-Linné-Weg 10, 50829, Cologne, Germany

    Marcus Garzón & Andreas Bachmair

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  1. Peter Schlögelhofer
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Correspondence to Andreas Bachmair.

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Schlögelhofer, P., Garzón, M., Kerzendorfer, C. et al. Expression of the ubiquitin variant ubR48 decreases proteolytic activity in Arabidopsis and induces cell death. Planta 223, 684–697 (2006). https://doi.org/10.1007/s00425-005-0121-z

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