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J. Biol. Chem., Vol. 275, Issue 41, 32227-32233, October 13, 2000
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From the Department of Pharmacology and Toxicology and
Comprehensive Cancer Center, Schools of Medicine and Dentistry,
University of Alabama at Birmingham, Birmingham, Alabama 35294
Treatment of human promyelocytic leukemia cells
(HL-60) with phorbol 12-myristate 13-acetate (PMA) is known to decrease
c-myc mRNA by blocking transcription elongation at
sites near the first exon/intron border. Treatment of HL-60 cells with
either PMA or bryostatin 1, which acutely activates protein
kinase C (PKC), decreased the levels of myc mRNA and
Myc protein. The inhibition of Myc synthesis accounted for the drop in
Myc protein, because PMA treatment had no effect on Myc turnover.
Treatment with PMA or bryostatin 1 increased nuclear protein binding to
MIE1, a c-myc intron 1 element that defines an RFX1-binding
X box. RFX1 antiserum supershifted MIE1-protein complexes. Increased
MIE1 binding was independent of protein synthesis and abolished by a
selective PKC inhibitor, which also prevented the effect of PMA on
myc mRNA and protein levels and Myc synthesis. PMA
treatment increased RFX1 in the nuclear fraction and decreased it in
the cytosol without affecting total RFX1. Transfection of HL-60 cells
with myc reporter gene constructs showed that the
RFX1-binding X box was required for the down-regulation of reporter
gene expression by PMA. These findings suggest that nuclear
translocation and binding of RFX1 to the X box cause the
down-regulation of myc expression, which follows acute PKC
activation in undifferentiated HL-60 cells.
Activation of Protein Kinase C Induces Nuclear Translocation of
RFX1 and Down-regulates c-myc via an Intron 1 X Box in
Undifferentiated Leukemia HL-60 Cells*
*
This work was supported by Grant GM60383 from the National
Institutes of Health.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Dept. of Pharmacology
and Toxicology, Volker Hall G133E, 1670 University Blvd., UAB,
Birmingham, AL 35294-0019. Tel.: 205-934-7434; Fax: 205-975-5841; E-mail: jeff.smith@ccc.uab.edu.
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