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(Received for publication, October 20, 1995, and in revised form, February 27, 1996)
From the Department of Cell Biology, Institute of Development,
Aging and Cancer, Tohoku University, Seiryomachi 4-1, Aoba-ku,
Sendai 980-77, Japan
The far upstream region (
1.2-0.9 kilobase
pairs) of the mouse glycophorin gene contains the locus control region
(LCR)-like region, which acts as an erythroid-specific enhancer
dependent on chromosomal integration in murine erythroleukemia (MEL)
cells. In the present study, we demonstrated that this region binds six
nuclear factors. The binding of GATA-1 to corresponding sites did not
show any change before or after induction with dimethyl sulfoxide, but
the binding of Spi-1/PU.l and an unidentified factor called glycophorin
regulatory element binding factor (GRBF) showed a change during
induction. While binding activity of Spi-l/PU.l dropped soon after
induction, the GRBF activity increased after induction when expression
of the glycophorin gene began. After identification of the consensus
binding site of GRBF, we cloned cDNA for that factor by
Southwestern method, and it was identified as a previously reported
transcription factor, delta, a murine form of YY-l which is a versatile
transcription factor. Mutation analysis in the delta/YY-1 binding site
within the LCR-like region indicated that delta/YY-1 acts as a
regulatory protein in combination with the E-box-binding protein that
binds to the neighboring sequence.
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