Isolation and functional analysis of the Catharanthus roseus deacetylvindoline-4-O-acetyltransferase gene promoter.
Madagascar periwinkle (Catharanthus roseus) produces many therapeutically valuable terpenoid indole alkaloids (TIAs), such as vinblastine and vincristine derived from the monomers vindoline and catharanthine. Deacetylvindoline-4-O-acetyltransferase (DAT) is a key enzyme for the terminal step of vindoline biosynthesis. In this research, the DAT gene promoter was cloned, sequenced, and analyzed. An approximately 1,773 bp genomic DNA fragment of DAT promoter was obtained. Sequence analysis revealed that DAT promoter contained several potential regulatory elements which were involved in the regulation of gene expression. To investigate its function, the promoter fragments with 5' deletions and gain-of-function deletions were fused to GUS reporter gene, and their expressions were analyzed by transient expression in C. roseus cell suspensions. The regulatory activity of DAT promoter was identified with fluorescence quantitative assays. Three TGACG motifs and one inverted motif (CGTCA) between -808 and -1,086 bp in the DAT promoter were found to be involved in methyljasmonate signal transduction pathway.
Wang
Quan
Q
Plant Biotechnology Research Center, Fudan-SJTU-Nottingham Plant Biotechnology R&D Center, School of Agriculture and Biology, Shanghai Jiao Tong University, 200240 Shanghai, China.
Yuan
Fang
F
Pan
Qifang
Q
Li
Meiya
M
Wang
Guofeng
G
Zhao
Jingya
J
Tang
Kexuan
K
D000085:Acetates / Q000494:pharmacology; D000123:Acetyltransferases / Q000235:genetics*; D001483:Base Sequence; D030008:Catharanthus / Q000201:enzymology / Q000235:genetics*; D003001:Cloning, Molecular; D003517:Cyclopentanes / Q000494:pharmacology; D018744:DNA, Plant / Q000235:genetics; D018506:Gene Expression Regulation, Plant; D017930:Genes, Reporter; D008969:Molecular Sequence Data; D054883:Oxylipins / Q000494:pharmacology; D011401:Promoter Regions, Genetic; D017422:Sequence Analysis, DNA; D024363:Transcription Initiation Site
D000085:Acetates; D003517:Cyclopentanes; D018744:DNA, Plant; D054883:Oxylipins; C072239:methyl jasmonate; D000123:Acetyltransferases; C064442:acetylcoenzyme A-deacetylvindoline 4-O-acetyltransferase