J. Biochem, 1995, Vol. 118, No. 3 593-600
© 1995 Japanese Biochemical Society
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Inducible Expression of Erythroid-Specific Mouse Glycophorin Gene Is Regulated by Proximal Elements and Locus Control Region-Like Sequence1
Department of Cell Biology, Institute of Development, Aging and Cancer, Tohoku University 4-1 Seiryomachi, Aoba-ku, Sendai 980
2To whom correspondence should be addressed. Tel: +81-22-274-1111 (ext. 3461), Fax: +81-22-272-5081
Cis-acting elements of the gene for mouse glycophorin, an erythroid-specific membrane glycoprotein, were determined by transient and stable transfection assays using murine erythroleukemia (MEL)cells. Cis-acting elements proximal to the transcription start site of the gene can be separated into the basal promoter (-1 to -91 bp)and the distal element (-133 to -92). The basal promoter contained GGTGG and GATA motifs and the distal element contained GATA-1 and NF-E2 motifs. Deletion analysis of the distal GATA site and its neighboring sequence and DNase-1 footprinting/EMSA (electrophoretic mobility shift assay) analysis indicated that induced nuclear factor binding to GATA-1 and its neighboring sequence may be required for expression during MEL cell differentiation induced by dimethyl sulfoxide treatment. The NF-E2 site was also shown to be essential for the promoter activity. An approximately 400 bp far upstream region (-1325 to -948bp) containing the binding motifs for GGGTGG, GATA-1 and NF-E2 showed no enhancing activity when this region was examined by transient transfection assay, but it did show enhancement of the differentiation-specific promoter activity in the stable transfection assay. The far upstream region of mouse glycophorin gene may have a function similar to that of the locus control region (LCR) of human ß -globin gene cluster.
1This work was supported by a Grant-in-Aid for Cancer Research from the Ministry of Education, Science and Culture of Japan.
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