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J. Biol. Chem., Vol. 278, Issue 25, 22928-22938, June 20, 2003

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Identification of a Human Homologue of the DREF Transcription Factor with a Potential Role in Regulation of the Histone H1 Gene^*

Nobuko Ohshima  , Masahide Takahashi  and Fumiko Hirose  ¶

From the Division of Biochemistry, Aichi Cancer Center Research Institute, Chikusa-ku, Nagoya 464-8681 and Department of Pathology, Nagoya Graduate School of Medicine, Showa-ku, Nagoya 466-8550, Japan

A human homologue (hDREF/KIAA0785) of Drosophila DREF, a transcriptional regulatory factor required for expression of genes involved in DNA replication and cell proliferation, was identified by BLAST search. Amino acid sequences corresponding to three regions highly conserved between two Drosophila species also proved to be very similar in the hDREF/KIAA0785 polypeptide. A consensus binding sequence (5'-TGTCG(C/T)GA(C/T)A) for hDREF/KIAA0785, determined by the CASTing method, overlapped with that for the Drosophila DREF (5'-TGTCGATA). We found hDREF/KIAA0785 binding sequences in the promoter regions of human genes related to cell proliferation. Analyses using a specific antibody revealed that an hDREF/KIAA0785 binds to the promoter region of the histone H1 gene. Co-transfection experiments with an hDREF/KIAA0785-expressing plasmid and a histone H1 promoter-directed luciferase reporter plasmid in HeLa cells revealed possible activation of the histone H1 promoter. Immunohistochemical analysis demonstrated that hDREF/KIAA0785 is localized in the nuclei. Although the expression level of the factor was found to be low in serum-deprived human normal fibroblasts, the amount was increased by adding serum to cultures and reached a maximum during S phase. RNA interference experiments targeting hDREF/KIAA0785 resulted in inhibition of S phase entry and reduction of histone H1 mRNA in HeLa cells. These results suggest that expression of hDREF/KIAA0785 may have a role in regulation of human genes related to cell proliferation.

Received for publication, March 26, 2003

^* This work was supported in part by grants-in-aid from the Ministry of Education, Science, Sports, Culture and Technology of Japan. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

^¶ To whom correspondence should be addressed. Tel.: 81-52-762-6111 (ext. 7220); Fax: 81-52-763-5233; E-mail: fsegawa{at}aichi-cc.jp.

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