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Originally published In Press as doi:10.1074/jbc.M006585200 on September 6, 2000

J. Biol. Chem., Vol. 275, Issue 50, 39411-39419, December 15, 2000
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RET Finger Protein Is a Transcriptional Repressor and Interacts with Enhancer of Polycomb That Has Dual Transcriptional Functions*

Yohei ShimonoDagger §, Hideki MurakamiDagger , Yoshinori Hasegawa§, and Masahide TakahashiDagger

From the Departments of Dagger  Pathology and § Internal Medicine I, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan

RET finger protein (RFP) belongs to the large B-box RING finger protein family and is known to become oncogenic by fusion with RET tyrosine kinase. Although RFP is reported to be a nuclear protein that is present in the nuclear matrix, its function is largely unknown. Here we show that RFP interacts with Enhancer of Polycomb (EPC) and strongly represses the gene transcription. Yeast two-hybrid assays revealed that the coiled-coil domain of RFP was associated with the EPcA domain and the carboxyl-terminal region of EPC. In addition, both proteins were co-precipitated from the lysates of human cells and mostly colocalized in the nucleus. Using the luciferase reporter-gene assay, we found that they repress the gene transcription activity independent of the differences of enhancers and promoters used, although the repressive activity of RFP was much stronger than that of EPC. The coiled-coil domain of RFP and the carboxyl-terminal region of EPC were most important for the repressive activity of each protein, whereas the EPcA domain had the transcription activating ability that is unique as the Polycomb group protein function. These results suggested that RFP may be involved in the epigenetic gene silencing mechanism cooperating with Polycomb group proteins and that EPC is a unique molecule with both repressive and transactivating activities.

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

The nucleotide sequence(s) reported in this paper for the human Enhancer of Polycomb gene (EPC) has been deposited in the GenBankTM/EBI Data Bank with accession number AF277374.

To whom correspondence should be addressed: Dept. of Pathology, Nagoya University School of Medicine, 65 Tsurumai-cho, Showa-ku, Nagoya 466-8550, Japan. Tel.: 81-52-744-2093; Fax: 81-52-744-2098; E-mail: mtakaha@med.nagoya-u.ac.jp.

Copyright © 2000 by The American Society for Biochemistry and Molecular Biology, Inc.
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