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GSyellow, a multifaceted tag for functional protein analysis in monocot and dicot plants

Nienke Besbrugge (UGent) , Jelle Van Leene (UGent) , Dominique Eeckhout (UGent) , Bernard Cannoot (UGent) , Shubhada Rajabhau Kulkarni (UGent) , Nancy De Winne (UGent) , Geert Persiau (UGent) , Eveline Van De Slijke (UGent) , Michiel Bontinck (UGent) , Stijn Aesaert (UGent) , et al.
(2018) PLANT PHYSIOLOGY. 177(2). p.447-464
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Abstract
The ability to tag proteins has boosted the emergence of generic molecular methods for protein functional analysis. Fluorescent protein tags are used to visualize protein localization, and affinity tags enable the mapping of molecular interactions by, for example, tandem affinity purification or chromatin immunoprecipitation. To apply these widely used molecular techniques on a single transgenic plant line, we developed a multifunctional tandem affinity purification tag, named GS(yellow), which combines the streptavidin-binding peptide tag with citrine yellow fluorescent protein. We demonstrated the versatility of the GS(yellow) tag in the dicot Arabidopsis (Arabidopsis thaliana) using a set of benchmark proteins. For proof of concept in monocots, we assessed the localization and dynamic interaction profile of the leaf growth regulator ANGUSTIFOLIA3 (AN3), fused to the GS(yellow) tag, along the growth zone of the maize (Zea mays) leaf. To further explore the function of ZmAN3, we mapped its DNA-binding landscape in the growth zone of the maize leaf through chromatin immunoprecipitation sequencing. Comparison with AN3 target genes mapped in the developing maize tassel or in Arabidopsis cell cultures revealed strong conservation of AN3 target genes between different maize tissues and across monocots and dicots, respectively. In conclusion, the GS(yellow) tag offers a powerful molecular tool for distinct types of protein functional analyses in dicots and monocots. As this approach involves transforming a single construct, it is likely to accelerate both basic and translational plant research.
Keywords
TANDEM AFFINITY PURIFICATION, CHROMATIN REMODELING COMPLEXES, CELL-SUSPENSION CULTURES, FLUORESCENT PROTEIN, TRANSCRIPTION FACTORS, ARABIDOPSIS-THALIANA, BUILDING-BLOCKS, LEAF GROWTH, MAIZE, GENOMICS

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Chicago
Besbrugge, Nienke, Jelle Van Leene, Dominique Eeckhout, Bernard Cannoot, Shubhada Rajabhau Kulkarni, Nancy De Winne, Geert Persiau, et al. 2018. “GSyellow, a Multifaceted Tag for Functional Protein Analysis in Monocot and Dicot Plants.” Plant Physiology 177 (2): 447–464.
APA
Besbrugge, N., Van Leene, J., Eeckhout, D., Cannoot, B., Kulkarni, S. R., De Winne, N., Persiau, G., et al. (2018). GSyellow, a multifaceted tag for functional protein analysis in monocot and dicot plants. PLANT PHYSIOLOGY, 177(2), 447–464.
Vancouver
1.
Besbrugge N, Van Leene J, Eeckhout D, Cannoot B, Kulkarni SR, De Winne N, et al. GSyellow, a multifaceted tag for functional protein analysis in monocot and dicot plants. PLANT PHYSIOLOGY. 2018;177(2):447–64.
MLA
Besbrugge, Nienke, Jelle Van Leene, Dominique Eeckhout, et al. “GSyellow, a Multifaceted Tag for Functional Protein Analysis in Monocot and Dicot Plants.” PLANT PHYSIOLOGY 177.2 (2018): 447–464. Print.
@article{8569528,
  abstract     = {The ability to tag proteins has boosted the emergence of generic molecular methods for protein functional analysis. Fluorescent protein tags are used to visualize protein localization, and affinity tags enable the mapping of molecular interactions by, for example, tandem affinity purification or chromatin immunoprecipitation. To apply these widely used molecular techniques on a single transgenic plant line, we developed a multifunctional tandem affinity purification tag, named GS(yellow), which combines the streptavidin-binding peptide tag with citrine yellow fluorescent protein. We demonstrated the versatility of the GS(yellow) tag in the dicot Arabidopsis (Arabidopsis thaliana) using a set of benchmark proteins. For proof of concept in monocots, we assessed the localization and dynamic interaction profile of the leaf growth regulator ANGUSTIFOLIA3 (AN3), fused to the GS(yellow) tag, along the growth zone of the maize (Zea mays) leaf. To further explore the function of ZmAN3, we mapped its DNA-binding landscape in the growth zone of the maize leaf through chromatin immunoprecipitation sequencing. Comparison with AN3 target genes mapped in the developing maize tassel or in Arabidopsis cell cultures revealed strong conservation of AN3 target genes between different maize tissues and across monocots and dicots, respectively. In conclusion, the GS(yellow) tag offers a powerful molecular tool for distinct types of protein functional analyses in dicots and monocots. As this approach involves transforming a single construct, it is likely to accelerate both basic and translational plant research.},
  author       = {Besbrugge, Nienke and Van Leene, Jelle and Eeckhout, Dominique and Cannoot, Bernard and Kulkarni, Shubhada Rajabhau and De Winne, Nancy and Persiau, Geert and Van De Slijke, Eveline and Bontinck, Michiel and Aesaert, Stijn and Impens, Francis and Gevaert, Kris and Van Damme, Dani{\"e}l and Van Lijsebettens, Maria and Inz{\'e}, Dirk and Vandepoele, Klaas and Nelissen, Hilde and De Jaeger, Geert},
  issn         = {0032-0889},
  journal      = {PLANT PHYSIOLOGY},
  keyword      = {TANDEM AFFINITY PURIFICATION,CHROMATIN REMODELING COMPLEXES,CELL-SUSPENSION CULTURES,FLUORESCENT PROTEIN,TRANSCRIPTION FACTORS,ARABIDOPSIS-THALIANA,BUILDING-BLOCKS,LEAF GROWTH,MAIZE,GENOMICS},
  language     = {eng},
  number       = {2},
  pages        = {447--464},
  title        = {GSyellow, a multifaceted tag for functional protein analysis in monocot and dicot plants},
  url          = {http://dx.doi.org/10.1104/pp.18.00175},
  volume       = {177},
  year         = {2018},
}

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