A plant DNA-binding protein increases the number of active preinitiation complexes in a human in vitro transcription system

Fumiaki Katagiri, Ken Ichi Yamazaki, Masami Horikoshi, Robert G. Roeder, Nam Hai Chua

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

Abstract

TGA1a is a tobacco DNA-binding protein that binds to the activation sequence-1 (as-1) element of the cauliflower mosaic virus 35S promoter. We have produced TGA1a in Escherichia coli, purified it from bacterial extracts, and examined its effect on transcription in a human in vitro system. Addition of TGA1a stimulates transcription by up to 20 times, and the stimulation is dependent on the presence of the as-1 element in the promoter. When transcription reinitiation is inhibited by 0.3 M KC1, activation is similar. Therefore, TGA1a activates transcription by increasing the number of active preinitiation complexes. After formation of the preinitiation complexes in the presence of TGA1a, oligonucleotides containing TGA1a-binding sites do not significantly affect the stimulated level of transcription. This result indicates that a complex remains committed to the promoter site after initiation and that this complex is used repeatedly during several initiation events. Our study demonstrates for the first time that a plant factor can activate transcription in a human in vitro system and that the activation mechanism of the plant factor is similar to that of mammalian factors.

Original languageEnglish (US)
Pages (from-to)1899-1909
Number of pages11
JournalGenes and Development
Volume4
Issue number11
DOIs
StatePublished - 1990

Keywords

  • ASF-1
  • As-1
  • RNA polymerase II transcription
  • TGA1a
  • Transcription activation mechanism

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