GSyellow, a Multifaceted Tag for Functional Protein Analysis in Monocot and Dicot Plants

被引：16

作者：

Besbrugge, Nienke ^{[1
,2
]}

Van Leene, Jelle ^{[1
,2
]}

Eeckhout, Dominique ^{[1
,2
]}

Cannoot, Bernard ^{[1
,2
]}

Kulkarni, Shubhada R. ^{[1
,2
,3
]}

De Winne, Nancy ^{[1
,2
]}

Persiau, Geert ^{[1
,2
]}

Van De Slijke, Eveline ^{[1
,2
]}

Bontinck, Michiel ^{[1
,2
]}

Aesaert, Stijn ^{[1
]}

Impens, Francis ^{[4
,5
,6
]}

Gevaert, Kris ^{[4
,5
]}

Van Damme, Daniel ^{[1
,2
]}

Van Lijsebettens, Mieke ^{[1
,2
]}

Inze, Dirk ^{[1
,2
]}

Vandepoele, Klaas ^{[1
,2
,3
]}

Nelissen, Hilde ^{[1
,2
]}

De Jaeger, Geert ^{[1
,2
]}

机构：

[1] Univ Ghent, Dept Plant Biotechnol & Bioinformat, B-9052 Ghent, Belgium

[2] VIB Ctr Plant Syst Biol, B-9052 Ghent, Belgium

[3] Univ Ghent, Bioinformat Inst Ghent, B-9052 Ghent, Belgium

[4] Univ Ghent, Dept Biochem, B-9000 Ghent, Belgium

[5] VIB Ctr Med Biotechnol, B-9000 Ghent, Belgium

[6] VIB Prote Core, B-9000 Ghent, Belgium

来源：

PLANT PHYSIOLOGY | 2018年 / 177卷 / 02期

基金：

欧洲研究理事会;

关键词：

TANDEM AFFINITY PURIFICATION; CHROMATIN REMODELING COMPLEXES; CELL-SUSPENSION CULTURES; FLUORESCENT PROTEIN; TRANSCRIPTION FACTORS; ARABIDOPSIS-THALIANA; BUILDING-BLOCKS; LEAF GROWTH; MAIZE; GENOMICS;

D O I：

10.1104/pp.18.00175

中图分类号：

Q94 [植物学];

学科分类号：

071001 ;

摘要：

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.

引用

页码：447 / 464

页数：18

共 62 条

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