J. Biochem, 1997, Vol. 121, No. 2 251-263
© 1997 Japanese Biochemical Society
research-article |
Conserved Structure, Regulatory Elements, and Transcriptional Regulation from the GATA-1 Gene Testis Promoter1
,2
*Department of Biochemistry, Tohoku University School of Medicine 2-1 Seiryomachi, Aoba-ku, Sendai 980-77
Institute of Basic Medical Sciences and Center for TARA, University of Tsukuba 1-1-1 Tennodai, Tsukuba 305
Department of Pediatrics, Hirosaki University School of Medicine 5 Zaifucho, Hirosaki 036
§Department of Biochemistry, Molecular Biology and Cell Biology, Northwestern University Evanston, IL 60208-3500, USA
||Unité 273 de l'Institut National de la Santé et de la Recherche Médicale Université de Nice 06108 Nice, Cedex 2, France
2To whom correspondence should be addressed. Tel: +81-298-53-3111, FAX: +81-298-53-6965 E-mail: masiya{at}igaku.md.tsukuba.ac.jp
Transcription factor GATA-1 was first identified in erythroid cells, but was later shown to also be expressed in Sertoli cells of the mouse testis. GATA-1 transcription in testis initiates from a different first exon (exon IT) than the erythroid mRNA (transcribed from exon IE). To begin to address the question of how expression of GATA-1 might be differentially regulated in Sertoli and erythroid cells, we have cloned and determined the structure of the IT promoters of both the rat and mouse GATA-1 genes. The transcription regulatory mechanism(s) controlling the synthesis of exon IT-derived mRNA was investigated by transfection of wild-type and mutant reporter genes, with and without co-transfected GATA factor expression plasmids, into either fibroblasts or Sertoli cell lines. Two GATA binding sites in the IT promoter were found to be required for GATA factor-mediated activation in fibroblasts: GATA-IT-directed reporter gene expression was activated only after co-transfection with GATA-1, implying that transcriptional activation of GATA-1 in the testis might be at least partially mediated through these GATA regulatory elements. We also found that the endogenous GATA-1 gene was silent in primary culture and two different Sertoli cell lines, and that the repression of co-transfected GATA-IT reporter genes could not be relieved by forced expression of GATA-1 in Sertoli cells. Thus the GATA-IT promoter may be under the control of a regulatory network in Sertoli cells which involves both positive and negative regulation of transcription, and conserved GATA motifs found in the IT promoter may berequired for transducing these effects.
1This work was supported in part by Grants-in-Aid from the Ministry of Education, Science, Sports and Culture of Japan (to MY), a grant from the Naito Foundation and Uehara Memorial Foundation (to MY), a grant from the Japanese Foundation of Promotion of Sciences (to MY and JDE), and a research grant from the NIH (GM28896 to JDE). KY was a Research Fellow of the Japanese Society for the Promotion of Science, whose present address is at the Institute of Microbial Disease, Osaka University.
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