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Retinoblastoma Binding Factor 1 Site in the Core Promoter Region of the Human RB Gene Is Activated by hGABP/E4TF1¹

Department of Preventive Medicine, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602 [Y. So., T. F., T. M., Y. O., D. K., T. S.]; Department of Oncology, The Institute of Medical Science, The University of Tokyo, 4-6-1 Shirokanedai, Minato-ku, Tokyo 108 [Y. Sh., J.-i. I.]; Department of Orthopedic Surgery, Wakayama Medical College, 7-Bacho, Wakayama 640 [T. M.]; Faculty of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuda-cho, Midori-ku, Yokohama 227 [J-i. S., M. G., H. W., H. H.], Japan

We previously reported two oncogenic point mutations present in the RB (retinoblastoma) gene promoter region, found at consensus Sp1 and ATF sites, respectively, and in two separate hereditary RB families. However, Sp1 protein was shown not to bind to the Sp1 site; this indicated that the Sp1 consensus site mutation was blocking the action of an alternative transcription factor, which we called RBF-1 (retinoblastoma binding factor 1). Subsequent purification of RBF-1 revealed it to be hGABP/E4TF1, a transactivator from the adenovirus early-region 4 promoter. In this study, we directly examined the effects of hGABP/E4TF1 on transactivation of the RB gene promoter through the RBF-1 site. As expected, hGABP/E4TF1 enhanced the core RB promoter activity, whereas it did not stimulate a mutant RBF-1 site. We therefore conclude that the most essential transcription factor in the human RB gene is likely to be hGABP/E4TF1.

¹ Supported in part by the Smoking Research Foundation Grant for Biomedical Research and the Ministry of Education, Science, Sports and Culture of Japan.

² Present address: Chugai Research Institute for Molecular Medicine, 153-2 Nagai, Niihari, Ibaraki 300-41, Japan.

³ To whom requests for reprints should be addressed, at Department of Preventive Medicine, Kyoto Prefectural University of Medicine, Kawaramachi-Hirokoji, Kamigyo-ku, Kyoto 602, Japan. Phone: 81-75-251-5339; Fax: 81-75-241-0792; E-mail: tsakai@basic.kpu-m.ac.jp.

Received 4/22/97. Accepted 6/16/97.

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