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Nucleic Acids Research, 2001, Vol. 29, No. 5 1228-1237
© 2001 Oxford University Press

Regulation of c-maf gene expression by Pax6 in cultured cells

Masaharu Sakai*, Mohamed S. Serria, Hiromi Ikeda, Kazuhiko Yoshida1, Junko Imaki2 and Shinzo Nishi

Department of Biochemistry and 1Department of Ophthalmology, Hokkaido University School of Medicine, N15, W7, Kita-ku, Sapporo 060-8638, Japan and 2Department of Anatomy, Nippon Medical School, 1-1-5, Sendagi, Bunkyou-ku, Tokyo 113-8602, Japan

c-Maf is a bZip transcription factor expressed in developmental and cellular differentiation processes. Recently, a c-maf knockout mouse model, showing abnormal lens development, has been reported. In order to study the regulation mechanisms of c-maf gene expression during the differentiation process we have cloned and functionally characterized the rat c-maf (maf-2) gene. The rat c-maf gene is an intronless gene, covering a length of 3.5 kb. Transient transfection analysis of the 5'-flanking region of the c-maf gene using luciferase as the reporter gene shows that Pax6, a master transcription factor for lens development, strongly activates the c-maf promoter construct. Endogenous c-maf is also activated by the Pax6 expression vector. Electrophoresis mobility shift assay and DNase I footprinting analysis show that at least three Pax6-binding sites are located in the 5'-flanking and 5'-non-coding regions of the rat c-maf gene. The c-maf gene was also markedly activated by its own product, c-Maf, through the MARE (Maf recognition element), suggesting that a positive autoregulatory mechanism controls this gene. In situ hybridization histochemical detection of Pax6 and c-Maf in the E14 lens showed that both mRNAs are expressed in the lens equator where lens epithelial cells are differentiating to lens fiber cells. These results suggest that a Pax6/c-Maf transcription factor cascade is working in lens development.

* To whom correspondence should be addressed. Tel: +81 11 706 5043; Fax: +81 11 706 7865; Email: m_sakai{at}med.hokudai.ac.jp


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