Wheat Dof transcription factor WPBF interacts with TaQM and activates transcription of an alpha-gliadin gene during wheat seed development.
Wheat prolamin-box binding factor (WPBF), a DOF transcription factor previously was isolated from wheat endosperm and suggested to function as an activator of prolamin gene expression during seed development. In this study, we showed that WPBF is expressed in all wheat tissues analyzed, and a protein, TaQM, was identified from a wheat root cDNA library, to interact with the Dof domain of WPBF. The specific interaction between WPBF and TaQM was confirmed by pull-down assay and bimolecular fluorescence complementation (BiFC) experiment. The expression patterns of TaQM gene are similar with that of WPBF. The GST-WPBF expressed in bacteria binds the Prolamin box (PB) 5'-TGTAAAG-3', derived from the promoter region of a native alpha-gliadin gene encoding a storage protein. Transient expression experiments in co-transfected BY-2 protoplast cells demonstrated that WPBF trans-activated transcription from native alpha-gliadin promoter through binding to the intact PB. When WPBF and TaQM are co-transfected together the transcription activity of alpha-gliadin gene was six-fold higher than when WPBF was transfected alone. Furthermore, the promoter activities of WPBF gene were observed in the seeds and the vascular system of transgenic Arabidopsis, which was identical to the expression profiles of WPBF in wheat. Hence, we proposed that WPBF functions not only during wheat seed development but also during other growth and development processes.
Dong
Guoqing
G
Department of Plant Genetics & Breeding and State Key Laboratory for Agrobiotechnology, China Agricultural University, Haidian district, Beijing, 100094, China.
Ni
Zhongfu
Z
Yao
Yingyin
Y
Nie
Xiuling
X
Sun
Qixin
Q
D024202:Electrophoretic Mobility Shift Assay; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D005903:Gliadin / Q000235:genetics*; D005966:Glucuronidase / Q000235:genetics / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000235:genetics / Q000378:metabolism; D010940:Plant Proteins / Q000235:genetics* / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012639:Seeds / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D014158:Transcription, Genetic; D014908:Triticum / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D020798:Two-Hybrid System Techniques
D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D005903:Gliadin; D005966:Glucuronidase