Human zinc-fingers and homeoboxes (ZHX) 1, a transcriptional repressor, was originally cloned as an interacting protein with the activation domain of the A subunit of nuclear factor-Y (NF-YA). As the first step in investigating the mechanism by which ZHX1 acts as a transcriptional repressor, we conducted a search of ZHX1-interacting proteins using a yeast two-hybrid system. Nuclear proteins such as ZHX1, transcriptional co-factors and DNA-binding proteins, zyxin, androgen-induced aldose reductase and eleven-nineteen lysine-rich leukaemia gene, as well as some unknown proteins, were cloned. Molecular cloning and determination of the nucleotide sequence of the full-length cDNA encoding a novel protein revealed that it consists of 956 amino acid residues and contains two zinc-finger (Znf) motifs and five homeodomains (HDs) as well as ZHX1. We concluded that the protein forms the ZHX family with ZHX1 and denoted it ZHX3. ZHX3 not only dimerizes with both ZHX1 and ZHX3, but also interacts with the activation domain of the NF-YA. Further analysis revealed that ZHX3 is a ubiquitous transcriptional repressor that is localized in nuclei and functions as a dimer. Lastly, the dimerization domain, the interaction domain with NF-YA, and the repressor domain are mapped to a region including the HD1 region, and two nuclear localization signals are mapped to the N-terminal through Znf1 and the HD2 region, respectively.
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Research Article|
July 01 2003
Analysis of zinc-fingers and homeoboxes (ZHX)-1-interacting proteins: molecular cloning and characterization of a member of the ZHX family, ZHX3
Kazuya YAMADA;
Kazuya YAMADA
1
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
1To whom correspondence should be addressed (e-mail kazuya@fmsrsa.fukui-med.ac.jp).
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Hiroko KAWATA;
Hiroko KAWATA
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
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Zhangfei SHOU;
Zhangfei SHOU
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
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Satoko HIRANO;
Satoko HIRANO
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
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Tetsuya MIZUTANI;
Tetsuya MIZUTANI
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
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Takashi YAZAWA;
Takashi YAZAWA
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
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Toshio SEKIGUCHI;
Toshio SEKIGUCHI
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
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Miki YOSHINO;
Miki YOSHINO
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
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Takashi KAJITANI;
Takashi KAJITANI
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
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Kaoru MIYAMOTO
Kaoru MIYAMOTO
∗Department of Biochemistry, Fukui Medical University, 23-3 Shimoaizuki, Matsuoka-cho, Fukui 910-1193, Japan
†CREST, Japan Science and Technology, Fukui 910-1193, Japan
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Publisher: Portland Press Ltd
Received:
December 02 2002
Revision Received:
February 28 2003
Accepted:
March 27 2003
Accepted Manuscript online:
March 27 2003
Online ISSN: 1470-8728
Print ISSN: 0264-6021
The Biochemical Society, London ©2003
2003
Biochem J (2003) 373 (1): 167–178.
Article history
Received:
December 02 2002
Revision Received:
February 28 2003
Accepted:
March 27 2003
Accepted Manuscript online:
March 27 2003
Citation
Kazuya YAMADA, Hiroko KAWATA, Zhangfei SHOU, Satoko HIRANO, Tetsuya MIZUTANI, Takashi YAZAWA, Toshio SEKIGUCHI, Miki YOSHINO, Takashi KAJITANI, Kaoru MIYAMOTO; Analysis of zinc-fingers and homeoboxes (ZHX)-1-interacting proteins: molecular cloning and characterization of a member of the ZHX family, ZHX3. Biochem J 1 July 2003; 373 (1): 167–178. doi: https://doi.org/10.1042/bj20021866
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