Modulation of the intracellular stability and toxicity of diphtheria toxin through degradation by the N-end rule pathway

The EMBO Journal
P O Falnes, S Olsnes

Abstract

The enzymatically active A-fragment of diphtheria toxin enters the cytosol of sensitive cells where it inhibits protein synthesis by inactivating elongation factor 2 (EF-2). We have constructed a number of diphtheria toxin mutants that are degraded by the N-end rule pathway in Vero cells, and that display a wide range of intracellular stabilities. The degradation could be inhibited by the proteasome inhibitor lactacystin, indicating that the proteasome is responsible for N-end rule-mediated degradation in mammalian cells. Previously, the N-end rule has been investigated by studying the co-translational degradation of intracellularly expressed beta-galactosidase. Our work shows that a mature protein entering the cytosol from the exterior can also be degraded by the N-end rule pathway with a similar, but not identical specificity to that previously found. We found a correlation between the intracellular stability of the mutants and their toxic effect on cells, thus demonstrating a novel manner of modulating the toxicity of a protein toxin. The data also indicate that the inactivation of EF-2 is the rate-limiting step in the intoxication process.

References

Feb 15, 1991·Proceedings of the National Academy of Sciences of the United States of America·R T Baker, A Varshavsky
Nov 15, 1991·European Journal of Biochemistry·K G KrieglsteinE Habermann
Nov 29, 1991·Science·J W TobiasA Varshavsky
Mar 24, 1989·Cell·A Bachmair, A Varshavsky
Jul 1, 1986·Proceedings of the National Academy of Sciences of the United States of America·A NicosiaR Rappuoli
Oct 18, 1988·Biochemistry·S M Arfin, R A Bradshaw
Jul 1, 1987·Proceedings of the National Academy of Sciences of the United States of America·S B CalderwoodJ J Mekalanos
Apr 15, 1985·European Journal of Biochemistry·F I LambJ M Lord
Jan 1, 1982·Proceedings of the National Academy of Sciences of the United States of America·M YamaizumiY Okada
Dec 1, 1980·The Journal of Cell Biology·K Sandvig, S Olsnes
Apr 25, 1995·Proceedings of the National Academy of Sciences of the United States of America·A Varshavsky
May 19, 1995·The Journal of Biological Chemistry·R T Baker, A Varshavsky
May 14, 1996·Proceedings of the National Academy of Sciences of the United States of America·F LévyA Varshavsky
Jan 1, 1995·Cold Spring Harbor Symposia on Quantitative Biology·A Varshavsky
Oct 29, 1996·Proceedings of the National Academy of Sciences of the United States of America·A Varshavsky
Dec 9, 1996·FEBS Letters·M Huguet SolerW Voelter

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Citations

Jun 30, 2000·Current Opinion in Cell Biology·P O Falnes, K Sandvig
Apr 22, 2009·Infection and Immunity·Juliette Morlon-GuyotBruno Beaumelle
Sep 5, 2008·PloS One·Pradeep K GuptaStephen H Leppla
May 14, 2010·PloS One·Yi ZhangDavid Morton Waisman
Nov 10, 2011·Toxins·Assaf Shapira, Itai Benhar
Aug 26, 2014·Toxins·Philip A Gurnev, Ekaterina M Nestorovich
Jan 26, 2016·ACS Central Science·Amy E Rabideau, Bradley L Pentelute
Jun 3, 2011·Protein Science : a Publication of the Protein Society·Alexander Varshavsky
Feb 27, 2004·Biochemical and Biophysical Research Communications·Byung-Moon KimYounghoon Lee
Nov 12, 2016·Toxins·Marta BorowiecKrzysztof Ginalski
Jan 26, 2005·Molecular Therapy : the Journal of the American Society of Gene Therapy·Sergey O TcherniukMaxim Y Balakirev
Jun 23, 2006·Genesis : the Journal of Genetics and Development·Damian BrockschniederDieter Riethmacher
Apr 5, 2000·The Journal of Biological Chemistry·T A Klink, R T Raines

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