PMID: 11607348Jan 15, 1993Paper

Use of a reporter transgene to generate arabidopsis mutants in ubiquitin-dependent protein degradation

Proceedings of the National Academy of Sciences of the United States of America
A BachmairJ Schell

Abstract

Ubiquitin-dependent proteolysis is a major proteolytic pathway in the cytoplasm and nucleus of eukaryotic cells. We introduced a gene encoding a substrate for this pathway into the genome of Arabidopsis thaliana. The transgene codes for a hybrid protein consisting of dihydrofolate reductase (DHFR, EC 1.5.1.3) fused to a degradation signal that is specifically recognized by components of the ubiquitin-dependent proteolysis pathway. Elevated concentrations of the DHFR protein confer resistance to the drug methotrexate, but rapid degradation prevents accumulation of the protein in the plant. Therefore, transgenic A. thaliana lines expressing the DHFR fusion protein are methotrexate-sensitive. Selection for mutants resistant to methotrexate resulted in plants impaired in degradation of the DHFR model substrate, as shown by an increase in protein level in the mutants.

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Citations

Jan 20, 1997·Biotechnology and Bioengineering·K S BatemanM A Anderson
Oct 1, 1996·Plant Molecular Biology·R D Vierstra
Oct 9, 2001·Trends in Plant Science·A BachmairF Eisenhaber
Jul 22, 2009·Proceedings of the National Academy of Sciences of the United States of America·Emmanuelle GracietFrank Wellmer
Sep 21, 2004·Annual Review of Plant Biology·Jan Smalle, Richard D Vierstra
Jul 8, 1998·Proceedings of the National Academy of Sciences of the United States of America·T PotuschakA Bachmair
Apr 9, 2010·Molecular Microbiology·D A DouganK Zeth
Dec 17, 2009·The Plant Journal : for Cell and Molecular Biology·Emmanuelle GracietFrank Wellmer
Jul 23, 2016·Nature Communications·Frederik FadenNico Dissmeyer
Oct 23, 2019·Journal of Integrative Plant Biology·Michael John HoldsworthAgata Zubrycka
Jul 9, 2014·Biological Chemistry·Frederik FadenNico Dissmeyer
Jan 1, 2020·Plant Direct·Christopher J TillMichael J Holdsworth
Feb 21, 2019·Annual Review of Plant Biology·A Harvey MillarWaltraud X Schulze
Aug 19, 2014·Journal of Proteome Research·Petra MajovskyWolfgang Hoehenwarter

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