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A Functional Composite Cis-Element for NFB and RBPJ in the Rat Pregnancy-Specific Glycoprotein Gene¹

^a Institute of Biological Chemistry, Academia Sinica, Nankang, Taipei 115, Taiwan ^b Graduate Institute of Biochemical Sciences, National Taiwan University, Taipei 106, Taiwan

ABSTRACT

The rat pregnancy-specific glycoprotein gene rnCGM3 is primarily expressed in the placenta. Previously, three DNase I footprinting sites (FPI, FPII, and FPIII) were identified in the rnCGM3 promoter region, a yeast one-hybrid screen was performed to identify the nuclear factors binding to the FPIII (5'-GCCTGGGAAAAAACTC-3') element, and RBPJ, a downstream effector of the Notch signaling pathway, was identified as one of the FPIII-binding factors. In the present study, the NFB member p65 was identified as another FPIII-binding factor. Electrophoretic mobility shift assays showed that NFB members, including p50 and p65, bound to the FPIII site. The core binding sequence in the FPIII element for p50 and p65 is GGGAAA, which overlaps with that for RBPJ. Competition exists between p50 and RBPJ for binding to the FPIII element. Transient expression analyses revealed that p65 significantly stimulated the expression of a reporter gene directed by the NFB core sequence in the FPIII element. However, RBPJ could block this stimulation. These results suggest that the regulation of rnCGM3 expression involves both NFB and RBPJ, and they are mutually exclusive in the FPIII element.

FOOTNOTES

First decision: 26 January 2001.

¹ This work was supported by grants (to H.C.) from the National Science Council (89-2311-B-001-229) and Academia Sinica of Taiwan.

² Correspondence. FAX: 011 886 2 27889759; hwchen{at}gate.sinica.edu.tw

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