Biochemical and redox characterization of the mediator complex and its associated transcription factor GeBPL, a GLABROUS1 enhancer binding protein. Shaikhali, J., Davoine, C., Brannstrom, K., Rouhier, N., Bygdell, J., Bjorklund, S., & Wingsle, G. Biochem J, 468(3):385–400, June, 2015. Edition: 2015/04/17
Biochemical and redox characterization of the mediator complex and its associated transcription factor GeBPL, a GLABROUS1 enhancer binding protein [link]Paper  doi  abstract   bibtex   
The eukaryotic mediator integrates regulatory signals from promoter-bound transcription factors (TFs) and transmits them to RNA polymerase II (Pol II) machinery. Although redox signalling is important in adjusting plant metabolism and development, nothing is known about a possible redox regulation of mediator. In the present study, using pull-down and yeast two-hybrid assays, we demonstrate the association of mediator (MED) subunits MED10a, MED28 and MED32 with the GLABROUS1 (GL1) enhancer-binding protein-like (GeBPL), a plant-specific TF that binds a promoter containing cryptochrome 1 response element 2 (CryR2) element. All the corresponding recombinant proteins form various types of covalent oligomers linked by intermolecular disulfide bonds that are reduced in vitro by the thioredoxin (TRX) and/or glutathione/glutaredoxin (GRX) systems. The presence of recombinant MED10a, MED28 and MED32 subunits or changes of its redox state affect the DNA-binding capacity of GeBPL suggesting that redox-driven conformational changes might modulate its activity. Overall, these results advance our understanding of how redox signalling affects transcription and identify mediator as a novel actor in redox signalling pathways, relaying or integrating redox changes in combination with specific TFs as GeBPL.
@article{shaikhali_biochemical_2015,
	title = {Biochemical and redox characterization of the mediator complex and its associated transcription factor {GeBPL}, a {GLABROUS1} enhancer binding protein},
	volume = {468},
	issn = {1470-8728 (Electronic) 0264-6021 (Linking)},
	url = {https://www.ncbi.nlm.nih.gov/pubmed/25877331},
	doi = {10.1042/BJ20150132},
	abstract = {The eukaryotic mediator integrates regulatory signals from promoter-bound transcription factors (TFs) and transmits them to RNA polymerase II (Pol II) machinery. Although redox signalling is important in adjusting plant metabolism and development, nothing is known about a possible redox regulation of mediator. In the present study, using pull-down and yeast two-hybrid assays, we demonstrate the association of mediator (MED) subunits MED10a, MED28 and MED32 with the GLABROUS1 (GL1) enhancer-binding protein-like (GeBPL), a plant-specific TF that binds a promoter containing cryptochrome 1 response element 2 (CryR2) element. All the corresponding recombinant proteins form various types of covalent oligomers linked by intermolecular disulfide bonds that are reduced in vitro by the thioredoxin (TRX) and/or glutathione/glutaredoxin (GRX) systems. The presence of recombinant MED10a, MED28 and MED32 subunits or changes of its redox state affect the DNA-binding capacity of GeBPL suggesting that redox-driven conformational changes might modulate its activity. Overall, these results advance our understanding of how redox signalling affects transcription and identify mediator as a novel actor in redox signalling pathways, relaying or integrating redox changes in combination with specific TFs as GeBPL.},
	language = {en},
	number = {3},
	urldate = {2021-06-07},
	journal = {Biochem J},
	author = {Shaikhali, J. and Davoine, C. and Brannstrom, K. and Rouhier, N. and Bygdell, J. and Bjorklund, S. and Wingsle, G.},
	month = jun,
	year = {2015},
	note = {Edition: 2015/04/17},
	keywords = {Arabidopsis Proteins/genetics/*metabolism, Arabidopsis thaliana, Arabidopsis/*metabolism, CryR2, DNA-Binding Proteins/genetics/*metabolism, GLABROUS1 enhancer-binding protein (GeBP), Glutaredoxins/metabolism, Glutathione/metabolism, Mediator Complex/genetics/*metabolism, Mutagenesis, Site-Directed, Mutation, Oxidation-Reduction, Promoter Regions, Genetic, Protein Subunits/genetics/*metabolism, Recombinant Proteins/metabolism, Response Elements, Thioredoxins/metabolism, Transcription Factors, Two-Hybrid System Techniques, deoxyribonucleic acid (DNA)-binding, mediator, redox},
	pages = {385--400},
}

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