Molecular Cloning of cDNA for a Novel Pea Dof Protein, PsDof1, and Its DNA-Binding Activity to the Promoter of PsDof1 Gene

Hikaru SEKI; Naoki NAKAMURA; Mizuri MARUTANI; Takeharu OKABE; Shiroh SANEMATSU; Yoshishige INAGAKI; Kazuhiro TOYODA; Tomonori SHIRAISHI; Tetsuji YAMADA; Yuki ICHINOSE

doi:10.5511/plantbiotechnology.19.251

Original Paper

Molecular Cloning of cDNA for a Novel Pea Dof Protein, PsDof1, and Its DNA-Binding Activity to the Promoter of PsDof1 Gene

Hikaru SEKI, Naoki NAKAMURA, Mizuri MARUTANI, Takeharu OKABE, Shiroh SANEMATSU, Yoshishige INAGAKI, Kazuhiro TOYODA, Tomonori SHIRAISHI, Tetsuji YAMADA, Yuki ICHINOSE

Author information

Hikaru SEKI
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Naoki NAKAMURA
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Mizuri MARUTANI
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Takeharu OKABE
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Shiroh SANEMATSU
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Yoshishige INAGAKI
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Kazuhiro TOYODA
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Tomonori SHIRAISHI
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Tetsuji YAMADA
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University
Yuki ICHINOSE
Laboratory of Plant Pathology & Genetic Engineering, Faculty of Agriculture, Okayama University

Keywords: Autoregulation, DNA-binding protein, Dof protein, Pisum sativum, Promoter, Transcription factor, Zinc-finger protein

JOURNAL FREE ACCESS

2002 Volume 19 Issue 4 Pages 251-260

DOI https://doi.org/10.5511/plantbiotechnology.19.251

Details

Abstract

A cDNA clone, designated PsDof1, encoding a novel member of the Dof DNA-binding protein family was isolated from pea. Random-binding-site selection and gel retardation experiments showed that the GST-PsDof1 recombinant protein bound to specific DNA sequences with an AAAG core. A metal-chelating agent, 1, 10-phenanthroline, abolished binding of GST-PsDof1 to DNA, but the addition of Zn²⁺ restored it, indicating that zinc ions are required for its DNA-binding activity. Interestingly, there are possible sites for binding PsDof1 in its promoter sequence. Indeed, GST-PsDof1 binds to the AAAG-containing promoter sequence of the PsDof1 gene with higher affinity. These results suggest that PsDof1 is a novel transcription factor that is involved in its own transcriptional regulation.

Corresponding author

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