Details

Autor(en) / Beteiligte

• Yan, Z H
• Medvedev, A
• Hirose, T
• Gotoh, H
• Jetten, A M

Titel

Characterization of the Response Element and DNA Binding Properties of the Nuclear Orphan Receptor Germ Cell Nuclear Factor/Retinoid Receptor-related Testis-associated Receptor

Ist Teil von

• The Journal of biological chemistry, 1997-04, Vol.272 (16), p.10565-10572

Ort / Verlag

United States: American Society for Biochemistry and Molecular Biology

Erscheinungsjahr

1997

Quelle

Free E-Journal (出版社公開部分のみ）

Beschreibungen/Notizen

• Recently, we have reported the cloning of the germ cell-specific, nuclear orphan receptor germ cell nuclear factor (GCNF)/RTR. In this study, we characterize the RTR response elements by an electrophoretic mobility shift assay/polymerase chain reaction-based, DNA binding site selection strategy. RTR binds with the greatest affinity to response elements containing TCA(AG(G/T)TCA) 2 (consensus RTR response element; conRTRE), to which it binds as a homodimer. RTR is also able to bind as a monomer to a single core motif TCAAG(G/T)TCA, albeit with a lower affinity. Mutation analysis supports the specific requirements of the 5′-flanking sequence and the core motif of the RTRE for optimal binding of RTR. An RTR-specific antiserum (RTR-Ab2) was raised that causes supershift of the RTR-conRTRE complex in EMSA. Based on the sequence of the conRTRE, we located a putative RTRE, referred to as P2-RE, in the 5′ promoter-flanking region of the mouse protamine 2 gene, which is induced during the same stage of spermatogenesis as RTR. The ability of RTR-Ab2 to cause a supershift of an RTR-RTRE complex with nuclear extracts from different tissues correlated with the tissue- and development-specific expression of RTR. Transfection of RTR in CV-1 cells was unable to cause RTRE-dependent transactivation of a CAT reporter gene; however, an RTR-VP16 fusion protein could induce transactivation through several RTREs, including P2-RE.