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Research article

Zinc-fingers and homeoboxes (ZHX) 2, a novel member of the ZHX family, functions as a transcriptional repressor

Hiroko KAWATA, Kazuya YAMADA, Zhangfei SHOU, Tetsuya MIZUTANI, Takashi YAZAWA, Miki YOSHINO, Toshio SEKIGUCHI, Takashi KAJITANI, Kaoru MIYAMOTO
Biochemical Journal Aug 01, 2003, 373 (3) 747-757; DOI: 10.1042/bj20030171
Hiroko KAWATA
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, Japan
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Kazuya YAMADA
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, JapanDepartment of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Zhangfei SHOU
Department of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Tetsuya MIZUTANI
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, JapanDepartment of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Takashi YAZAWA
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, JapanDepartment of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Miki YOSHINO
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, JapanDepartment of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Toshio SEKIGUCHI
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, JapanDepartment of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Takashi KAJITANI
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, JapanDepartment of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Kaoru MIYAMOTO
Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology (JST), Fukui 910-1193, JapanDepartment of Biochemistry, Fukui Medical University, Shimoaizuki, Matsuoka, Fukui 910-1193, Japan
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Abstract

Zinc-fingers and homeoboxes (ZHX) 1 is a transcription factor that interacts with the activation domain of the A subunit of nuclear factor-Y (NF-YA). Using a yeast two-hybrid system, a novel ubiquitous transcription factor ZHX2 as a ZHX1-interacting protein was cloned. ZHX2 consists of 837 amino acid residues and contains two zinc-finger motifs and five homeodomains (HDs) as well as ZHX1. The mRNA is expressed among various tissues. ZHX2 not only forms a heterodimer with ZHX1, but also forms a homodimer. Moreover, ZHX2 interacts with the activation domain of NF-YA. Further analysis revealed that ZHX2 is a transcriptional repressor that is localized in the nuclei. Since ZHX2 shares a number of properties in common with ZHX1, we conclude that all these come under the ZHX family. The minimal functional domains of ZHX2 were then characterized. The dimerization domain with both ZHX1 and ZHX2 is the region containing HD1, the domain that interacts with NF-YA is the HD1 to HD2 region, the repressor domain is the HD1 to a proline-rich region. Lastly, using an immunoprecipitation assay, we showed that ZHX2 intrinsically interacts with NF-YA in HEK-293 cells and that ZHX2 represses the promoter activity of the cdc25C gene stimulated by NF-Y in Drosophila Schneider line 2 cells. Thus the ZHX family of proteins may participate in the expression of a number of NF-Y-regulated genes via a more organized transcription network.

  • dimerization
  • nuclear factor-Y
  • nuclear localization signal
  • transcriptional repressor
  • zinc-fingers and homeoboxes 1 (ZHX1)

Footnotes

  • Nucleotide sequences of pAD-L26 and the entire coding of human ZHX2 cDNA have been submitted to the DNA Databank of Japan under the accession numbers AB081946 and AB083653 respectively.

  • Abbreviations used: AD, activation domain; DBD, DNA-binding domain; GFP, green fluorescent protein; GST, glutathione S-transferase; HD, homeodomain; NF-Y, nuclear factor-Y; NLS, nuclear localization signal; P/CAF, p300/cAMP-response-element-binding protein-associated factor; RT, reverse transcriptase; SL2, Schneider line 2; Sp, specificity protein; SV40, simian virus 40; ZHX, zinc-fingers and homeoboxes; Znf, zinc finger.

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August 2003

Volume: 373 Issue: 3

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Zinc-fingers and homeoboxes (ZHX) 2, a novel member of the ZHX family, functions as a transcriptional repressor
Hiroko KAWATA, Kazuya YAMADA, Zhangfei SHOU, Tetsuya MIZUTANI, Takashi YAZAWA, Miki YOSHINO, Toshio SEKIGUCHI, Takashi KAJITANI, Kaoru MIYAMOTO
Biochemical Journal Aug 2003, 373 (3) 747-757; DOI: 10.1042/bj20030171
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Zinc-fingers and homeoboxes (ZHX) 2, a novel member of the ZHX family, functions as a transcriptional repressor
Hiroko KAWATA, Kazuya YAMADA, Zhangfei SHOU, Tetsuya MIZUTANI, Takashi YAZAWA, Miki YOSHINO, Toshio SEKIGUCHI, Takashi KAJITANI, Kaoru MIYAMOTO
Biochemical Journal Aug 2003, 373 (3) 747-757; DOI: 10.1042/bj20030171

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Keywords

dimerization
nuclear factor-Y
nuclear localization signal
transcriptional repressor
zinc-fingers and homeoboxes 1 (ZHX1)

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