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The Arabidopsis iron-sulfur protein GRXS17 is a target of the ubiquitin E3 ligases RGLG3 and RGLG4

Astrid Nagels Durand (UGent) , Sabrina Iñigo (UGent) , Andrés Ritter Traub (UGent) , Elisa Iniesto, Rebecca De Clercq (UGent) , An Staes (UGent) , Jelle Van Leene (UGent) , Vicente Rubio, Kris Gevaert (UGent) , Geert De Jaeger (UGent) , et al.
(2016) PLANT AND CELL PHYSIOLOGY. 57(9). p.1801-1813
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Abstract
The stability of signaling proteins in eukaryotes is often controlled by post-translational modifiers. For polyubiquitination, specificity is assured by E3 ubiquitin ligases. Although plant genomes encode hundreds of E3 ligases, only few targets are known, even in the model Arabidopsis thaliana. Here, we identified the monothiol glutaredoxin GRXS17 as a substrate of the Arabidopsis E3 ubiquitin ligases RING DOMAIN LIGASE 3 (RGLG3) and RGLG4 using a substrate trapping approach involving tandem affinity purification of RING-dead versions. Simultaneously, we used a ubiquitin-conjugating enzym (UBC) panel screen to pinpoint UBC30 as a cognate E2 UBC capable of interacting with RGLG3 and RGLG4 and mediating auto-ubiquitination of RGLG3 and ubiquitination of GRXS17 in vitro. Accordingly, GRXS17 is ubiquitinated and degraded in an RGLG3-and RGLG4-dependent manner in planta. The truncated hemoglobin GLB3 also interacted with RGLG3 and RGLG4 but appeared to obstruct RGLG3 ubiquitination activity rather than being its substrate. Our results suggest that the RGLG family is intimately linked to the essential element iron.
Keywords
STRESS-RESPONSE, MONOTHIOL GLUTAREDOXIN, RING, HEMOGLOBIN, IDENTIFICATION, CARDIAC-HYPERTROPHY, SACCHAROMYCES-CEREVISIAE, FUNCTIONAL-CHARACTERIZATION, GLUTAREDOXIN 3 PICOT, Ubiquitination, KeyWords Plus:TANDEM AFFINITY PURIFICATION, Thioredoxin, Post-translational modification, Glutaredoxin, Author Keywords:Arabidopsis, E3 ubiquitin ligase

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Chicago
Nagels Durand, Astrid, Sabrina Iñigo, Andrés Ritter Traub, Elisa Iniesto, Rebecca De Clercq, An Staes, Jelle Van Leene, et al. 2016. “The Arabidopsis Iron-sulfur Protein GRXS17 Is a Target of the Ubiquitin E3 Ligases RGLG3 and RGLG4.” Plant and Cell Physiology 57 (9): 1801–1813.
APA
Nagels Durand, A., Iñigo, S., Ritter Traub, A., Iniesto, E., De Clercq, R., Staes, A., Van Leene, J., et al. (2016). The Arabidopsis iron-sulfur protein GRXS17 is a target of the ubiquitin E3 ligases RGLG3 and RGLG4. PLANT AND CELL PHYSIOLOGY, 57(9), 1801–1813.
Vancouver
1.
Nagels Durand A, Iñigo S, Ritter Traub A, Iniesto E, De Clercq R, Staes A, et al. The Arabidopsis iron-sulfur protein GRXS17 is a target of the ubiquitin E3 ligases RGLG3 and RGLG4. PLANT AND CELL PHYSIOLOGY. 2016;57(9):1801–13.
MLA
Nagels Durand, Astrid, Sabrina Iñigo, Andrés Ritter Traub, et al. “The Arabidopsis Iron-sulfur Protein GRXS17 Is a Target of the Ubiquitin E3 Ligases RGLG3 and RGLG4.” PLANT AND CELL PHYSIOLOGY 57.9 (2016): 1801–1813. Print.
@article{8174326,
  abstract     = {The stability of signaling proteins in eukaryotes is often controlled by post-translational modifiers. For polyubiquitination, specificity is assured by E3 ubiquitin ligases. Although plant genomes encode hundreds of E3 ligases, only few targets are known, even in the model Arabidopsis thaliana. Here, we identified the monothiol glutaredoxin GRXS17 as a substrate of the Arabidopsis E3 ubiquitin ligases RING DOMAIN LIGASE 3 (RGLG3) and RGLG4 using a substrate trapping approach involving tandem affinity purification of RING-dead versions. Simultaneously, we used a ubiquitin-conjugating enzym (UBC) panel screen to pinpoint UBC30 as a cognate E2 UBC capable of interacting with RGLG3 and RGLG4 and mediating auto-ubiquitination of RGLG3 and ubiquitination of GRXS17 in vitro. Accordingly, GRXS17 is ubiquitinated and degraded in an RGLG3-and RGLG4-dependent manner in planta. The truncated hemoglobin GLB3 also interacted with RGLG3 and RGLG4 but appeared to obstruct RGLG3 ubiquitination activity rather than being its substrate. Our results suggest that the RGLG family is intimately linked to the essential element iron.},
  author       = {Nagels Durand, Astrid and I{\~n}igo, Sabrina and Ritter Traub, Andr{\'e}s and Iniesto, Elisa and De Clercq, Rebecca and Staes, An and Van Leene, Jelle and Rubio, Vicente and Gevaert, Kris and De Jaeger, Geert and Pauwels, Laurens and Goossens, Alain},
  issn         = {0032-0781},
  journal      = {PLANT AND CELL PHYSIOLOGY},
  language     = {eng},
  number       = {9},
  pages        = {1801--1813},
  title        = {The Arabidopsis iron-sulfur protein GRXS17 is a target of the ubiquitin E3 ligases RGLG3 and RGLG4},
  url          = {http://dx.doi.org/10.1093/pcp/pcw122},
  volume       = {57},
  year         = {2016},
}

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