The mode of expression and promoter analysis of the arcA gene, an auxin-regulated gene in tobacco BY-2 cells.
The arcA, a member of the G protein beta-subunit family, was isolated from tobacco BY-2 cells as an auxin-responsive gene. Characterization of arcA, which should help to elucidate the function of the gene product in the plant cells, was performed with emphasis on the mode of expression and the analysis of its promoter. Accumulation of the arcA message was detected only after treatments with auxins and not after treatments with other phytohormones or CdCl2, implying that responsiveness of arcA was exclusive to auxin. The putative arcA promoter region was fused to a reporter gene for beta-glucuronidase (GUS), and transient expression was analyzed in tobacco BY-2 cells. Two series of arcA promoter/GUS chimeric genes were constructed. One consisted of a set of 5' nested deletions of the arcA promoter connected to the gene for GUS and the other consisted of a variety of the arcA promoter fragments fused to a minimal promoter-GUS construct. The results indicated that the promoter sequence covering four sets of direct repeats (-562 to -167) was necessary for the sufficient response of arcA promoter to auxin in BY-2 cells. Moreover, irrespective of auxin treatment, elevated activity of GUS driven by this promoter fragment was detected, a result that implies that this region behaves an enhancer in BY-2 cells.
Ishida
S
S
Department of Biological Sciences, Graduate School of Science, University of Tokyo, Japan.
Takahashi
Y
Y
Nagata
T
T
D001483:Base Sequence; D002678:Chimera / Q000235:genetics; D003001:Cloning, Molecular; D018744:DNA, Plant / Q000235:genetics; D004742:Enhancer Elements, Genetic; D019204:GTP-Binding Proteins / Q000737:chemistry / Q000235:genetics; D015870:Gene Expression; D017343:Genes, Plant; D017930:Genes, Reporter; D005966:Glucuronidase / Q000235:genetics; D007210:Indoleacetic Acids / Q000378:metabolism*; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D011401:Promoter Regions, Genetic; D011487:Protein Conformation; D014026:Nicotiana / Q000235:genetics* / Q000378:metabolism*
D018744:DNA, Plant; D007210:Indoleacetic Acids; D005966:Glucuronidase; D019204:GTP-Binding Proteins