A highly sensitive plant hybrid protein assay system based on the Spm promoter and TnpA protein for detection and analysis of transcription activation domains

Michael Schläppi, Ramesh Raina, Nina Fedoroff

Research output: Contribution to journalArticle

8 Citations (Scopus)

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 tile 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.

Original languageEnglish (US)
Pages (from-to)717-725
Number of pages9
JournalPlant Molecular Biology
Volume32
Issue number4
StatePublished - Nov 1996
Externally publishedYes

Fingerprint

Plant Proteins
transcriptional activation
Transcription
Transcriptional Activation
Assays
Chemical activation
promoter regions
DNA-binding domains
DNA-Binding Proteins
assays
binding sites
Proteins
proteins
Binding Sites
Protein Multimerization
TATA box
TATA Box
DNA Transposable Elements
DNA-binding proteins
dimerization

Keywords

  • hybrid protein
  • plant assay
  • Spm
  • TnpA
  • transcription activation
  • transposon

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

A highly sensitive plant hybrid protein assay system based on the Spm promoter and TnpA protein for detection and analysis of transcription activation domains. / Schläppi, Michael; Raina, Ramesh; Fedoroff, Nina.

In: Plant Molecular Biology, Vol. 32, No. 4, 11.1996, p. 717-725.

Research output: Contribution to journalArticle

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