Journal Article

The Transcriptional Activation Domain of the Plant-Specific Dof1 Factor Functions in Plant, Animal, and Yeast Cells

Shuichi Yanagisawa

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 42, issue 8, pages 813-822
Published in print August 2001 | ISSN: 0032-0781
Published online August 2001 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pce105
The Transcriptional Activation Domain of the Plant-Specific Dof1 Factor Functions in Plant, Animal, and Yeast Cells

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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Maize Dof1, one of the plant-specific Dof transcription factors, is involved in light-regulated gene expression. To elucidate the molecular mechanism underlying the activity of Dof1, in vivo functional analyses were carried out using transient expression assays with maize mesophyll protoplasts. The results suggest that the Dof domain alone, the region conserved among Dof factors, can mediate interaction with DNA in vivo and distinct Dof1 activities in greening and etiolated protoplasts. A region rich in basic amino acids was identified as a nuclear localization signal. Deletion analysis defined the transcriptional activation domain of 48 amino acids located in the C-terminus of Dof1. This activation domain was also found to be functional in both human cells and yeast, implying that Dof1 may stimulate transcription through a mechanism evolutionarily conserved among eukaryotes. A computer homology search with known transcription factors revealed that the activation domain of Dof1 displayed only a limited similarity to Activation domain II of animal transcription factor GATA-4. Mutational analysis revealed the critical role of a tryptophan residue within the activation domain of Dof1, as had been shown in Activation domain II of GATA-4.

Keywords: Key words: DNA-binding domain — Dof transcription factor — GATA family — Light regulation — Nuclear localization signal — Transcriptional activation domain.; Abbreviations: BGH, bovine growth hormone polyadenylation signal; CAT, chloramphenicol acetyltransferase; CMV, cytomegalovirus immediate-early promoter; GAL4-AD, transcriptional activation domain of GAL4; GAL4-BD, DNA binding domain of GAL4; GAL4-BD-AD, protein with GAL4-BD fused to GAL4-AD; β-gal, β-galactosidase; GUS, β-glucuronidase; HSP, heat shock protein; NOS, nopaline synthase; NLS, nuclear localization signal; PCR, polymerase chain reaction; sGFP, synthetic green fluorescent protein; SV40, simian virus 40.

Journal Article.  7048 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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