A proximal promoter region of Arabidopsis DREB2C confers tissue-specific expression under heat stress.
The dehydration-responsive element-binding factor 2C (DREB2C) is a member of the CBF/DREB subfamily of proteins, which contains a single APETALA2/Ethylene responsive element-binding factor (AP2/ERF) domain. To identify the expression pattern of the DREB2C gene, which contains multiple transcription cis-regulatory elements in its promoter, an approximately 1.4 kb upstream DREB2C sequence was fused to the β-glucuronidase reporter gene (GUS) and the recombinant p1244 construct was transformed into Arabidopsis thaliana (L.) Heynh. The promoter of the gene directed prominent GUS activity in the vasculature in diverse young dividing tissues. Upon applying heat stress (HS), GUS staining was also enhanced in the vasculature of the growing tissues. Analysis of a series of 5'-deletions of the DREB2C promoter revealed that a proximal upstream sequence sufficient for the tissue-specific spatial and temporal induction of GUS expression by HS is localized in the promoter region between -204 and -34 bps relative to the transcriptional start site. Furthermore, electrophoretic mobility shift assay (EMSA) demonstrated that nuclear protein binding activities specific to a -120 to -32 bp promoter fragment increased after HS. These results indicate that the TATA-proximal region and some latent trans-acting factors may cooperate in HS-induced activation of the Arabidopsis DREB2C promoter.
Systems and Synthetic Agrobiotech Center and PMBBRC, Division of Applied Life Science, Gyeongsang National University, Jinju 660-701, Korea.
Huan
H
Chen
Jihyun
J
Je
Chieun
C
Song
Jung Eun
JE
Hwang
Chae Oh
CO
Lim
D017360:Arabidopsis / Q000235:genetics* / Q000502:physiology; D029681:Arabidopsis Proteins / Q000235:genetics* / Q000378:metabolism; D020029:Base Pairing / Q000235:genetics; D018744:DNA, Plant / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism; D024202:Electrophoretic Mobility Shift Assay; D018506:Gene Expression Regulation, Plant; D005966:Glucuronidase / Q000378:metabolism; D018869:Heat-Shock Response / Q000235:genetics; D006358:Hot Temperature; D009687:Nuclear Proteins / Q000378:metabolism; D009928:Organ Specificity / Q000235:genetics*; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic; D011485:Protein Binding / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D017384:Sequence Deletion / Q000235:genetics; D013312:Stress, Physiological / Q000235:genetics*; D016385:TATA Box / Q000235:genetics; D013997:Time Factors
D029681:Arabidopsis Proteins; D018744:DNA, Plant; D004268:DNA-Binding Proteins; C573537:DREB2C protein, Arabidopsis; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins; D005966:Glucuronidase