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Molecular and Cellular Biology, October 2004, p. 8556-8566, Vol. 24, No. 19
0270-7306/04/$08.00+0     DOI: 10.1128/MCB.24.19.8556-8566.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Subcellular Localization of RPB5-Mediating Protein and Its Putative Functional Partner

Luvsanjav Delgermaa, Naoyuki Hayashi, Dorjbal Dorjsuren, Takahiro Nomura, Le Thi-Thu Thuy, and Seishi Murakami^*

Department of Molecular Oncology, Cancer Research Institute, Kanazawa University, Kanazawa, Japan

Received 7 February 2004/ Returned for modification 9 March 2004/ Accepted 10 June 2004

We previously identified a novel cellular protein, RPB5-mediating protein (RMP), that retains corepressor activity and functionally antagonizes transcriptional modulation via hepatitis B virus X protein. The subcellular localization of RMP was examined using green fluorescent protein-fused protein forms. We found that a nuclear localization signal (NLS) and a coiled-coil (CC) domain functioning as a cytoplasmic localization signal (CLS) are important for the subcellular localization of RMP. The CLS apparently acts dominantly, since RMP was mostly localized in the cytoplasm with weak and diffuse signals in the nucleus, and the NLS was indispensable for the nuclear localization of RMP only in the absence of the CLS. Using a yeast two-hybrid method, we isolated a putative corepressor, DNA methyltransferase 1-associating protein (DMAP1), which was found to bind to the CC domain of RMP. DMAP1 facilitated the nuclear localization of RMP and the corepressor activity of RMP in a dose-dependent manner by interacting with the CC domain of RMP. These results are discussed in light of a recent paper showing a novel evolutionarily conserved role of URI in the TOR signaling pathway.

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* Corresponding author. Mailing address: Department of Molecular Oncology, Cancer Research Institute, Kanazawa University, Takara-machi 13-1, Kanazawa 920-0934, Japan. Phone: 81-76-265-2731. Fax: 81-76-234-4501. E-mail: semuraka{at}kenroku.kanazawa-u.ac.jp.

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Molecular and Cellular Biology, October 2004, p. 8556-8566, Vol. 24, No. 19
0022-538X/04/$08.00+0     DOI: 10.1128/MCB.24.19.8556-8566.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

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