Structural and functional analysis of rice catalase-B gene promoter: presence of Dof and CAAT binding site

• Published in: Plant molecular biology reporter Vol. 25; no. 1-2; pp. 71 - 82
• Main Authors: Mondal, P, Dey, N, Dash, A.K, Chatterjee, A, Sahu, B.B, Panda, B, Maiti, I.B, Sabat, S.C
• Format: Journal Article
• Language: English
• Published: New York Springer Nature B.V 01.06.2007
• Subjects: beta-glucuronidase; Binding sites; Catalase; Deoxyribonuclease; Deoxyribonucleic acid; DNA; DNA-binding domains; enzyme activity; Flaking; Functional analysis; Gene deletion; Gene expression; molecular sequence data; Nucleotide sequence; Oryza sativa; promoter regions; Regulatory sequences; reporter genes; rice; Structure-function relationships; transcription (genetics)
• ISSN: 0735-9640; 1572-9818
• DOI: 10.1007/s11105-007-0011-6

A 5' flaking nucleotide sequence of 1,101 bp of rice catalase-B gene (CatB) coding sequence having coordinates from -808 to +294 with respect to the transcription start site was transcriptionally fused to the β-glucuronidase gene (GUS), and functionally analyzed through transient and stable expression assay systems to identify the minimal promoter sequence containing the important cis-regulatory regions controlling its expression. The progressive up-stream and down-stream (5'/3') deletion analysis of the 1,101-bp fragment revealed that minimal nucleotide sequences spanning from -121 to +56 possessed maximum GUS activity, which was nearly six- to eightfold higher than GUS activity driven by CaMV35S promoter. DNase I footprint and gel retardation analysis of the minimal promoter sequence with nuclear extract revealed the presence of a "AAAAG" nucleotide signature referring to DNA-binding with one finger (Dof) transcription factor. In addition, the presence of CAAT-box sequence was also evident. The Dof interacting sequence in CatB promoter of rice has been established for the first time in this work.