Document Type
Article
Language
eng
Format of Original
9 p.
Publication Date
11-1996
Publisher
Springer
Source Publication
Plant Molecular Biology
Source ISSN
0167-4412
Abstract
TnpA is a multifunctional DNA binding protein encoded by the maize Suppressor-mutator (Spm) transposable element. TnpA is required for transposition and is a repressor of the unmethylated Spm promoter. While analyzing protein domains using a yeast GAL4-based hybrid system in transiently transformed tobacco cells, we found that TnpA represses the >10-fold transcriptional activation observed when the GAL4 DNA-binding domain is used alone. By contrast, compared to the backgroundless TnpA DNA-binding domain alone, 33- to 45-fold activation of the Spm promoter was observed when the VP16 activation domain was fused to it. TnpA-binding sites, but no TATA box, were required for transcription activation. Among the TnpA deletion derivatives tested, those retaining the coding sequences for the DNA-binding and protein dimerization domains gave the highest level of transcription activation when fused with the VP16 activation domain. The TnpA gene and TnpA-binding sites in the short Spm promoter therefore provide a novel, highly sensitive single-hybrid system for identifying and studying plant transcription activation domains in plant cells.
Recommended Citation
Schläppi, Michael; Raina, Ramesh; and Fedoroff, Nina V., "A Highly Sensitive Plant Hybrid Protein Assay System Based on the Spm Promoter and TnpA Protein for Detection and Analysis of Transcription Activation Domains" (1996). Biological Sciences Faculty Research and Publications. 38.
https://epublications.marquette.edu/bio_fac/38
Comments
Accepted version. Plant Molecular Biology, Vol. 32, No. 4 (November 1996): 717-725. DOI. © 1996 Springer. Used with permission.
Michael Schläppi was affiliated with the Carnegie Institution of Washington at the time of publication.